Compounds active at a novel site on receptor-operated calcium channels useful for treatment of neurological disorders and diseases

ABSTRACT

Method and compositions for treating a patient having a neurological disease or disorder, such as stroke, head trauma, spinal cord injury, spinal cord ischemia, ischemia- or hypoxia-induced nerve cell damage, epilepsy, anxiety, neuropsychiatric or cognitive deficits due to ischemia or hypoxia such as those that frequently occur as a consequence of cardiac surgery under cardiopulmonary bypass, or neurodegenerative diseases such as Alzheimer&#39;s Disease, Huntington&#39;s Disease, Parkinson&#39;s Disease, or amyotrophic lateral sclerosis (ALS).

This application is a continuation of co-pending U.S. application Ser.No. 09/825,373 filed on Apr. 2, 2001, which is a continuation of U.S.application Ser. No. 09/186,341 filed Nov. 4, 1998, now U.S. Pat. No.6,211,245, which is a continuation of U.S. application Ser. No.08/873,011, filed Jun. 11, 1997, now abandoned, which is acontinuation-in-part of U.S. application Ser. No. 08/869,154, filed Jun.4, 1997, now abandoned, which is a continuation-in-part of U.S.application Ser. No. 08/763,480, filed Dec. 11, 1996, now U.S. Pat. No.6,017,965, which is a continuation-in-part of U.S. application Ser. No..08/663,013, filed Jun. 7, 1996, now abandoned, which is acontinuation-in-part of U.S. application Ser. No. 08/485,038, filed Jun.7, 1995, now U.S. Pat. No. 6,071,970, which was a National Stageapplication of International Patent Appl. No. PCT/US94/12293(WO95/21612), filed Oct. 26, 1994 and designating the United States,which is a continuation-in-part of U.S. application Ser. No. 08/288,668,filed Aug. 9, 1994, now abandoned, which is a continuation-in-part ofU.S. application Ser. No. 08/194,210, filed Feb. 8, 1994, now abandoned,which is a continuation-in-part of U.S. Ser. No. 08/014,813, filed Feb.8, 1993, now abandoned, each of which are hereby incorporated byreference herein in their entirety, including all Tables, Figures, andclaims.

FIELD OF THE INVENTION

This invention relates to compounds useful as neuroprotectants,anticonvulsants, anxiolytics, analgesics, muscle relaxants or adjuvantsto general anesthetics. The invention relates as well to methods usefulfor the treatment of neurological disorders and diseases, including, butnot limited to, global and focal ischemic and hemorrhagic stroke, headtrauma, spinal cord injury, hypoxia-induced nerve cell damage such as incardiac arrest or neonatal distress, epilepsy, anxiety, andneurodegenerative diseases such as Alzheimer's Disease, Huntington'sDisease, Parkinson's Disease, and amyotrophic lateral sclerosis (ALS).The invention relates as well to methods of screening for compoundsactive at a novel site on receptor-operated calcium channels, andthereby possessing therapeutic utility as neuroprotectants,anticonvulsants, anxiolytics, analgesics, muscle relaxants or adjuvantsto general anesthetics, and/or possessing potential therapeutic utilityfor the treatment of neurological disorders and diseases as describedabove.

BACKGROUND OF THE INVENTION

The following is a description of relevant art, none of which isadmitted to be prior art to the claims.

Glutamate is the major excitatory neurotransmitter in the mammalianbrain. Glutamate binds or interacts with one or more glutamate receptorswhich can be differentiated pharmacologically into several subtypes. Inthe mammalian central nervous system (CNS) there are three main subtypesof ionotropic glutamate receptors, defined pharmacologically by theselective agonists N-methyl-D-aspartate (NMDA), kainate (KA), andα-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA). The NMDAreceptor has been implicated in a variety of neurological pathologiesincluding stroke, head trauma, spinal cord injury, epilepsy, anxiety,and neurodegenerative diseases such as Alzheimer's Disease (Watkins andCollingridge, The NMDA Receptor, Oxford: IRL Press, 1989). A role forNMDA receptors in nociception and analgesia has been postulated as well(Dickenson, A cure for wind-up: NMDA receptor antagonists as potentialanalgesics. Trends Pharmacol. Sci. 11: 307, 1990). More recently, AMPAreceptors have been widely studied for their possible contributions tosuch neurological pathologies (Fisher and Bogousslavsky, Evolving towardeffective therapy for acute ischemic stroke. J. Amer. Med. Assoc. 270:360, 1993; Yamaguchi et al., Anticonvulsant activity of AMPA/kainateantagonists: Comparison of GYKI 52466 and NBQX in maximal electroshockand chemoconvulsant seizure models. Epilepsy Res. 15: 179, 1993).

When activated by glutamate, the endogenous neurotransmitter, the NMDAreceptor permits the influx of extracellular calcium (Ca²⁺) and sodium(Na⁺) through an associated ion channel. The NMDA receptor allowsconsiderably more influx of Ca²⁺ than do kainate or AMPA receptors (butsee below), and is an example of a receptor-operated Ca²⁺ channel.Normally, the channel is opened only briefly, allowing a localized andtransient increase in the concentration of intracellular Ca²⁺([Ca²⁺]_(i)), which, in turn, alters the functional activity of thecell. However, prolonged increases in [Ca²⁺]_(i), resulting from chronicstimulation of the NMDA receptor, are toxic to the cell and lead to celldeath. The chronic elevation in [Ca²⁺]_(i), resulting from stimulationof NMDA receptors, is said to be a primary cause of neuronaldegeneration following a stroke (Choi, Glutamate neurotoxicity anddiseases of the nervous system. Neuron 1: 623, 1988). Overstimulation ofNMDA receptors is also said to be involved in the pathogenesis of someforms of epilepsy (Dingledine et al., Excitatory amino acid receptors inepilepsy. Trends Pharmacol. Sci. 11: 334, 1990), anxiety (Wiley andBalster, Preclinical evaluation of N-methyl-D-aspartate antagonists forantianxiety effects: A review. In: Multiple Sigma and PCP ReceptorLigands: Mechanisms for Neuromodulation and Neuroprotection? NPP Books,Ann Arbor, Mich., pp. 801-815, 1992), neurodegenerative diseases(Meldrum and Garthwaite, Excitatory amino acid neurotoxicity andneurodegenerative disease. Trends Pharmacol. Sci. 11: 379, 1990), andhyperalgesic states (Dickenson, A cure for wind-up: NMDA receptorantagonists as potential analgesics. Trends Pharmacol. Sci. 11: 307,1990)

The activity of the NMDA receptor-ionophore complex is regulated by avariety of modulatory sites that can be targeted by selectiveantagonists. Competitive antagonists, such as the phosphonate AP5, actat the glutamate binding site, whereas noncompetitive antagonists, suchas phencyclidine (PCP), MK-801 or magnesium (Mg²⁺), act within theassociated ion channel (ionophore). There is also a glycine binding sitethat can be blocked selectively with compounds such as 7-chlorokynurenicacid. There is evidence suggesting that glycine acts as a co-agonist, sothat both glutamate and glycine are necessary to fully elicit NMDAreceptor-mediated responses. Other potential sites for modulation ofNMDA receptor function include a zinc (Zn²⁺) binding site and a sigmaligand binding site. Additionally, endogenous polyamines such asspermine are believed to bind to a specific site and so potentiate NMDAreceptor function (Ransom and Stec, Cooperative modulation of [³H]MK-801binding to the NMDA receptor-ion channel complex by glutamate glycineand polyamines. J. Neurochem. 51: 830, 1988). The potentiating effect ofpolyamines on NMDA receptor function may be mediated via a specificreceptor site for polyamines; polyamines demonstrating agonist,antagonist, and inverse agonist activity have been described (Reynolds,Arcaine is a competitive antagonist of the polyamine site on the NMDAreceptor. Europ. J. Pharmacol. 177: 215, 1990; Williams et al.,Characterization of polyamines having agonist, antagonist, and inverseagonist effects at the polyamine recognition site of the NMDA receptor.Neuron 5: 199, 1990). Radioligand binding studies have demonstratedadditionally that higher concentrations of polyamines inhibit NMDAreceptor function (Reynolds and Miller, Ifenprodil is a novel type ofNMDA receptor antagonist: Interaction with polyamines. Molec. Pharmacol.36: 758, 1989; Williams et al., Effects of polyamines on the binding of[³H]MK-801 to the NMDA receptor: Pharmacological evidence for theexistence of a polyamine recognition site. Molec. Pharmacol. 36: 575,1989; Sacaan and Johnson, Characterization of the stimulatory andinhibitory effects of polyamines on [³H]TCP binding to the NMDAreceptor-ionophore complex. Molec. Pharmacol. 37: 572, 1990). Thisinhibitory effect of polyamines on NMDA receptors is probably anonspecific effect (i.e., not mediated via the polyamine receptor)because patch clamp electro-physiological studies have demonstrated thatthis inhibition is produced by compounds previously shown to act at thepolyamine receptor as either agonists or antagonists (Donevan et al.,Arcaine Blocks N-Methyl-D-Aspartate Receptor Responses by an OpenChannel Mechanism: Whole-Cell and Single-Channel Recording Studies inCultured Hippocampal Neurons. Molec. Pharmacol. 41: 727, 1992; Rock andMacdonald, Spermine and Related Polyamines Produce a Voltage-DependentReduction of NMDA Receptor Single-Channel Conductance. Molec. Pharmacol.42: 157, 1992).

Recent studies have demonstrated the molecular diversity of glutamatereceptors (reviewed by Nakanishi, Molecular Diversity of GlutamateReceptors and Implications for Brain Function. Science 258: 597, 1992).At least five distinct NMDA receptor subunits (NMDAR1 and NMDAR2Athrough NMDAR2D), each encoded by a distinct gene, have been identifiedto date. Also, in NMDAR1, alternative splicing gives rise to at leastsix additional isoforms. It appears that NMDAR1 is a necessary subunit,and that combination of NMDAR1 with different members of NMDAR2 formsthe fully functional NMDA receptor-ionophore complex. The NMDAreceptor-ionophore complex, thus, can be defined as a hetero-oligomericstructure composed of at least NMDAR1 and NMDAR2 subunits; the existenceof additional, as yet undiscovered, subunits is not excluded by thisdefinition. NMDAR1 has been shown to possess binding sites forglutamate, glycine, Mg²⁺ MK-801, and Zn²⁺. The binding sites for sigmaligands and polyamines have not yet been localized on NMDA receptorsubunits, although ifenprodil recently has been reported to be morepotent at the NMDAR2B subunit than at the NMDAR2A subunit (Williams,Ifenprodil discriminates subtypes of the N-methyl-D-aspartate receptor:selectivity and mechanisms at recombinant heteromeric receptors. Mol.Pharmacol. 44: 851, 1993).

Several distinct subtypes of AMPA and kainate receptors have been clonedas well (reviewed by Nakanishi, Molecular diversity of glutamatereceptors and implications for brain function. Science 258: 597, 1992).Of particular relevance are the AMPA receptors designated GluR1, GluR2,GluR3, and GluR4 (also termed GluRA through GluRD), each of which existsin one of two forms, termed flip and flop, which arise by RNAalternative splicing. GluR1, GluR3 and GluR4, when expressed ashomomeric or heteromeric receptors, are permeable to Ca²⁺, and aretherefore examples of receptor-operated Ca²⁺ channels. Expression ofGluR2 alone or in combination with the other subunits gives rise to areceptor which is largely impermeable to Ca²⁺. As most native AMPAreceptors studied in situ are not Ca²⁺-permeable (discussed above), itis believed that such receptors in situ possess at least one GluR2subunit.

Furthermore, it is hypothesized that the GluR2 subunit is functionallydistinct by virtue of the fact that it contains an argihine residuewithin the putative pore-forming transmembrane region II; GluR1, GluR3and GluR4 all contain a glutamine residue in this critical region(termed the Q/R site, where Q and R are the single letter designationsfor glutamine and arginine, respectively). The activity of the AMPAreceptor is regulated by a number of modulatory sites that can betargeted by selective antagonists (Honore et al., Quinoxalinediones:potent competitive non-NMDA glutamate receptor antagonists. Science 241:701, 1988; Donevan and Rogawski, GYKI 52466, a 2,3-benzodiazepine, is ahighly selective, noncompetitive antagonist of AMPA/kainate receptorresponses. Neuron 10: 51, 1993). Competitive antagonists such as NBQXact at the glutamate binding site, whereas compounds such as GYKI 52466appear to act noncompetitively at an associated allosteric site.

Compounds that act as competitive or noncompetitive antagonists at theNMDA receptor are said to be effective in preventing neuronal cell deathin various in vitro neurotoxicity assays (Meldrum and Garthwaite,Excitatory amino acid neurotoxicity and neurodegenerative disease.Trends Pharmacol. Sci. 11: 379, 1990) and in in vivo models of stroke(Scatton, Theraceutic potential of NMDA receptor antagonists in ischemiccerebrovascular disease in Drug Strategies in the Prevention andTreatment of Stroke, IBC Technical Services Ltd., 1990). Such compoundsare also effective anticonvulsants (Meldrum, Excitatory amino acidneurotransmission in epilepsy and anticonvulsant therapy in ExcitatoryAmino Acids. Meldrum, Moroni, Simon, and Woods (Eds.), New York: RavenPress, p. 655, 1991), anxiolytics (Wiley and Balster, Preclinicalevaluation of N-methyl-D-aspartate antagonists for antianxiety effects:A review. In: Multiple Sigma and PCP Receptor Ligands: Mechanisms forNeuromodulation and Neuroprotection? NPP Books, Ann Arbor, Mich., pp.801-815, 1992), and analgesics (Dickenson, A cure for wind-up: NMDAreceptor antagonists as potential analgesics. Trends Pharmacol. Sci. 11:307, 1990), and certain NMDA receptor antagonists may lessen dementiaassociated with Alzheimer's Disease (Hughes, Merz' novel approach to thetreatment of dementia. Script No. 1666: 24, 1991).

Similarly, AMPA receptor antagonists have come under intense scrutiny aspotential therapeutic agents for the treatment of such neurologicaldisorders and diseases. AMPA receptor antagonists have been shown topossess neuroprotectant (Fisher and Bogousslavsky, Evolving towardeffective therapy for acute ischemic stroke. J. Amer. Med. Assoc. 270:360, 1993) and anticonvulsant (Yamaguchi et al., Anticonvulsant activityof AMPA/kainate antagonists: comparison of GYKI 52466 and NBQX inmaximal electroshock and chemoconvulsant seizure models. Epilepsy Res.15: 179, 1993) activity in animal models of ischemic stroke andepilepsy, respectively.

The nicotinic cholinergic receptor present in the mammalian CNS isanother example of a receptor-operated Ca²⁺, channel (Deneris et al.,Pharmacological and functional diversity of neuronal nicotinicacetylcholine receptors. Trends Pharmacol. Sci. 12: 34, 1991). Severaldistinct receptor subunits have been cloned, and these subunits can beexpressed, in Xenopus oocytes for example, to form functional receptorswith their associated cation channels. It is hypothesized that suchreceptor-ionophore complexes are heteropentameric structures. Thepossible role of nicotinic receptor-operated Ca²⁺ channels in thepathology of neurological disorders and diseases such as ischemicstroke, epilepsy and neurodegenerative diseases has been largelyunexplored.

It has been demonstrated previously that certain spider and wasp venomscontain arylalkylamine toxins (also called polyamine toxins, arylaminetoxins, acylpolyamine toxins or polyamine amide toxins) with activityagainst glutamate receptors in the mammalian CNS (for reviews seeJackson and Usherwood, Spider toxins as tools for dissecting elements ofexcitatory amino acid transmission. Trends Neurosci. 11: 278, 1988;Jackson and Parks, Spider Toxins: Recent Applications In Neurobiology.Annu. Rev. Neurosci. 12: 405, 1989; Saccomano et al., Polyamine spidertoxins: Unique pharmacological tools. Annu. Rep. Med. Chem. 24: 287,1989; Usherwood and Blagbrough, Spider Toxins Affecting GlutamateReceptors: Polyamines in Therapeutic Neurochemistry. Pharmacol. Therap.52: 245, 1991; Kawai, Neuroactive Toxins of Spider Venoms. J. Toxicol.Toxin Rev. 10: 131, 1991). Arylalkylamine toxins were initially reportedto be selective antagonists of the AMPA/kainate subtypes of glutamatereceptors in the mammalian CNS (Kawai et al., Effect of a spider toxinon glutaminergic synapses in the mammalian brain. Biomed. Res. 3: 353,1982; Saito et al., Spider Toxin (JSTX) blocks glutamate synapse inhippocampal pyramidal neurons. Brain Res. 346: 397, 1985; Saito et al.,Effects of a spider toxin (JSTX) on hippocampal CAl neurons in vitro.Brain Res. 481: 16, 1989; Akaike et al., Spider toxin blocks excitatoryamino acid responses in isolated hippocampal pyramidal neurons.Neurosci. Lett. 79: 326, 1987; Ashe et al., Argiotoxin-636 blocksexcitatory synaptic transmission in rat hippocampal CAl pyramidalneurons. Brain Res. 480: 234, 1989; Jones et al., Philanthotoxin blocksquisqualate-induced, AMPA-induced and kainate-induced, but notNMDA-induced excitation of rat brainstem neurones in vivo. Br. J.Pharmacol. 101: 968, 1990). Subsequent studies have demonstrated thatwhile certain arylalkylamine toxins are both nonpotent and nonselectiveat various glutamate receptors, other arylalkylamines are both verypotent and selective at antagonizing responses mediated by NMDA receptoractivation in the mammalian CNS (Mueller et al., Effects of polyaminespider toxins on NMDA receptor-mediated transmission in rat hippocampusin vitro. Soc. Neurosci. Abst. 15: 945, 1989; Mueller et al., Arylaminespider toxins antagonize NMDA receptor-mediated synaptic transmission inrat hippocampal slices. Synapse 9: 244, 1991; Parks et al., Polyaminespider toxins block NMDA receptor-mediated increases in cytosoliccalcium in cerebellar granule neurons. Soc. Neurosci. Abst. 15: 1169,1989; Parks et al., Arylamine toxins from funnel-web spider (Agelenopsisaperta) venom antagonize N-methyl-D-aspartate receptor function inmammalian brain. J. Biol. Chem. 266: 21523, 1991; Priestley et al.,Antagonism of responses to excitatory amino acids on rat corticalneurones by the spider toxin, argiotoxin-636. Br. J. Pharmacol. 97:1315, 1989; Draguhn et al., Argiotoxin-636 inhibits NMDA-activated ionchannels expressed in Xenopus oocytes. Neurosci. Lett. 132: 1.87, 1991;Kiskin et al., A highly potent and selective N-methyl-D-asparatereceptor antagonist from the venom of the Agelenopsis aperta spider.Neuroscience 51: 11, 1992; Brackley et al., Selective antagonism ofnative and cloned kainate and NMDA receptors by polyamine-containingtoxins. J. Pharmacol. Exptl. Therap. 266: 1573, 1993; Williams, Effectsof Agelenopsis aperta toxins on the N-methyl-D-aspartate receptor:Polyamine-like and high-affinity antagonist actions. J. Pharmacol.Exptl. Therap. 266: 231, 1993). Inhibition of nicotinic cholinergicreceptors by the arylalkylamine toxin philanthotoxin has also beenreported (Rozental et al., Allosteric inhibition of nicotinicacetylcholine receptors of vertebrates and insects by philanthotoxin. J.Pharmacol. Exptl. Therap. 249: 123, 1989).

Parks et al. (Arylamine toxins from funnel-web spider (Agelenopsisaperta) venom antagonize N-methyl-D-aspartate receptor function inmammalian brain. J. Biol. Chem. 266: 21523, 1991), describearylalkylamine spider toxins (α-agatoxins) which antagonize NMDAreceptor function in mammalian brain. The authors discuss the mechanismof action of arylalkylamine toxins, and indicate that an NMDAreceptor-operated ion channel is the probable site of action of theα-agatoxins, and most probably other spider venom arylalkylamines. Theystate:

-   -   The discovery that endogenous polyamines in the vertebrate brain        modulate the function of NMDA receptors suggests that the        arylamine toxins may produce their antagonism via a        polyamine-binding site on glutamate receptors. Brackley et al.        studied the effects of spermine and philanthotoxin 433 on the        responses evoked by application of excitatory amino acids in        Xenopus oocytes injected with mRNA from rat or chick brain.        These authors reported that, at concentrations below those that        antagonize glutamate receptor function, both spermine and        philanthotoxin potentiate the effects of excitatory amino acids        and some other neurotransmitters. On the basis of these and        other data, Brackley et al. concluded that the arylamine toxins        may, by binding nonspecifidally to the membranes of excitable        cells, reduce membrane fluidity and alter receptor function. The        validity of this intriguing idea for NMDA receptor function is        not well supported by two recent binding studies. Reynolds        reported that argiotoxin 636 inhibits the binding of [³H]MK-801        to rat brain membranes in a manner that is insensitive to        glutamate, glycine, or spermidine. This author concluded that        argiotoxin 636 exerts a novel inhibitory effect on the NMDA        receptor complex by binding to one of the Mg²⁺ sites located        within the NMDA-gated ion channel. Binding data reported by        Williams et al. also support the conclusion that argiotoxin 636        does not act primarily at the polyamine modulatory site on the        NMDA receptor, but rather acts directly to produce an        activity-dependent block of the ion channel. It is already known        that compounds such as phencyclidine and ketamine can block the        ion channels associated with both arthropod muscle glutamate        receptors and mammalian NMDA receptors. Thus, it seems possible        that vertebrate and invertebrate glutamate receptors share        additional binding sites for allosteric modulators of receptor        function, perhaps related to divalent cation-binding sites.        Clearly, considerable additional work will be needed to        determine if the arylamines define such a novel regulatory site.

Usherwood and Blagbrough (Spider Toxins Affecting Glutamate Receptors:Polyamines in Therapeutic Neurochemistry. Pharmacol. Therap. 52: 245,1991) describe a proposed intracellular binding site for arylalkylaminetoxins (polyamine amide toxins) located within the membrane potentialfield referred to as the QUIS-R channel selectivity filter. The authorspostulate that the binding site for polyamine amide toxins may occurclose to the internal entrance of the channel gated by the QUIS-R oflocust muscle. The authors also note that one such toxin,argiotoxin-636, selectively antagonizes the NMDA receptor in culturedrat cortical neurons.

Gullak et al. (CNS binding sites of the novel NMDA antagonist Arg-636.Soc. Neurosci. Abst. 15: 1168, 1989), describe argiotoxin-636 (Arg-636)as a polyamine (arylalkylamine) toxin component of a spider venom. Thistoxin is said to block NMDA-induced elevation of cGMP in anoncompetitive fashion. The authors state that:

-   -   [¹²⁵I]Arg-636 bound to rat forebrain membranes with K_(d) and        B_(max) values of 11.25 μM and 28.95 pmol/mg protein (80%        specific). The ability of other known polyamines and recently        discovered polyamines from Agelenopsis aperta to inhibit binding        paralleled neuroactivity as functional NMDA antagonists. No        other compounds tested were able to block specific binding.

The authors then stated that polyamines (arylalkylamines) may antagonizeresponses to NMDA by interacting with membrane ion channels.

Seymour and Mena (In vivo NMDA antagonist activity of the polyaminespider venom component, argiotoxin-636. Soc. Neurosci. Abst. 15: 1168,1989) describe studies that are said to show that argiotoxin-636 doesnot significantly affect locomotor activity at doses that are effectiveagainst audiogenic seizures in DBA/2 mice, and that it significantlyantagonizes NMDA-induced seizures with a minimal effective dose of 32mg/kg given subcutaneously (s.c.).

Herold and Yaksh (Anesthesia and muscle relaxation with intrathecalinjections of AR636 and AG489, two acylpolyamine. spider toxins, inrats. Anesthesiology 77: 507, 1992) describe studies that are said toshow that the arylalkylamine argiotoxin-636 (AR636), but notagatoxin-489 (AG489), produces muscle relaxation and anesthesiafollowing intrathecal administration in rats.

Williams (Effects of Agelenopsis aperta toxins on-theN-methyl-D-aspartate receptor:.Polyamine-like and high-affinityantagonist actions, J. Pharmacol. Exptl. Therap. 266: 231, 1993) reportsthat the α-agatoxins (arylalkylamines) Agel-489 and Agel-505 enhance thebinding of [³H]MK-801 to NMDA receptors on membranes prepared from ratbrain by an action at the stimulatory polyamine receptor; polyaminereceptor agonists occluded the stimulatory effects of Agel-489 andAgel-505 and polyamine receptor antagonists inhibited the stimulatoryeffect of Agel-505. Higher concentrations of Agel-489 and Agel-505, andargiotoxin-636 at all concentrations tested, had inhibitory effects onthe binding of [³H]MK-801. In Xenopus oocytes voltage-clamped at −70 mV,Agel-505 inhibited responses to NMDA with an IC₅₀, of 13 nM; this effectof Agel-505 occurred at concentrations approximately 10,000-fold lowerthan those that affected [³H]MK-801 binding. Responses to kainate wereinhibited only 11% by 30 nM Agel-505. The antagonism of NMDA-inducedcurrents by Agel-505 was strongly voltage-dependent, consistent with anopen-channel blocking effect of the toxin. Williams states:

-   -   Although α-agatoxins can interact at the positive allosteric        polyamine site on the NMDA receptor, stimulatory effects        produced by this interaction may be masked in functional assays        due to a separate action of the toxins as high-affinity,        noncompetitive antagonists of the receptor.

Brackley et al. (Selective antagonism of native and cloned kainate andNMDA receptors by polyamine-containing toxins, J. Pharmacol. Exp.Therap. 266: 1573, 1993) report that the polyamine-containing toxins(arylalkylamines) philanthotoxin-343 (PhTX-343) and argiotoxin-636(Arg-636) produce reversible, noncompetitive, partly voltage-dependentantagonism of kainate- and NMDA-induced currents in Xenopus oocytesinjected with rat brain RNA. Arg-636 was demonstrated to be selectivefor NMDA-induced responses (IC₅₀=0.04 μM) compared to kainate-inducedresponses (IC₅₀=0.07 μM), while PhTX-343 was selective forkainate-induced responses IC₅₀=0.12 μM) compared to NMDA-inducedresponses (IC₅₀=2.5 μM). Arg-636 more potently antagonized responses toNMDA in Xenopus oocytes expressing cloned NMDAR1 subunits (IC₅₀=0.09 μM)than responses to kainate in oocytes expressing either cloned GluR1(IC₅₀=3.4 μM) or GluR1+GluR2 subunits (IC₅₀=300 μM). PhTX-343, on theother hand, was equipotent at antagonizing NMDAR1 (IC₅₀=2.19 μM) andGluR1 (IC₅₀=2.8 μM) , but much less potent against GluR1+GluR2 subunits(IC₅₀=270 μM)

Raditsch et al. (Subunit-specific block of cloned NMDA receptors byargiotoxin-636. FEBS Lett. 324: 63, 1993) report that Arg-636 morepotently antagonizes responses in Xenopus oocytes expressingNMDAR1+NMDAR2A subunits (IC₅₀=9 nM) or NMDAR1+NMDAR2B subunits (IC₅₀=2.5nM) than NMDAR1+NMDAR2C subunits (IC₅₀=460 nM) even though all of thereceptor subunits contain an asparagine residue in the putativepore-forming transmembrane region II (the Q/R site, as discussed above).The authors state that the large difference in Arg-636 sensitivitybetween NMDAR1+NMDAR2A and NMDAR1+NMDAR2C channels “must be conferred byother structural determinants.”

Herlitz et al. (Argiotoxin detects molecular differences in AMPAreceptor-channels. Neuron 10: 1131, 1993) report that Arg-636antagonizes subtypes of AMPA receptors in a voltage- and use-dependentmanner consistent with open-channel blockade. Arg-636 potentlyantagonizes Ca²-permeable AMPA receptors comprised of GluRAi (K_(i)=0.35μM), GluRCi (K_(i)=0.23 μM), or GluRDi subunits (K_(i)=0.43 μM), whilebeing essentially ineffective against Ca², -impermeable GluRBi subunitsat concentrations up to 10 μM.

Other data reported by these investigators strongly suggest that the Q/Rsite in the putative pore-forming transmembrane region II is of primaryimportance in determining Arg-636 potency and Ca²⁺ permeability.

Blaschke et al. (A single amino acid determines the subunit-specificspider toxin block ofα-amino-3-hydroxy-5-methylisoxazole-4-propionate/kainate receptorchannels. Proc. Natl. Acad. Sci. USA 90: 6528, 1993) report that thearylalkylamine JSTX-3 potently antagonizes responses to kainate inXenopus oocytes expressing GluR1 (IC₅₀)=0.04 μM) or GluR3 (IC₅₀=0.03 μM)subunits, but that expressed receptors in which a GluR2 subunit ispresent are essentially unaffected by the toxin. Site-directedmutagenesis studies strongly implicate the Q/R site as the primary siteinfluencing toxin potency.

Nakanishi et al. (Bioorganic studies of transmitter receptors withphilanthotoxin analogs. Pure Appl. Chem., in press) have synthesized anumber of highly potent photoaffinity labeled philanthotoxin (PhTX)analogs. Such analogs have been studied on expressed nicotiniccholinergic receptors as a model system for receptor-operated calciumchannels. These investigators suggest that these PhTX analogs block theion channel with the hydrophobic headpiece of the toxin binding to asite near the cytoplasmic surface while the polyamine tail extends intothe ion channel from the cytoplasmic side.

SUMMARY OF THE INVENTION

Applicant has examined the structural diversity and biological activityof arylalkylamines (sometimes referred to as arylamine toxins, polyaminetoxins, acylpolyamine toxins or polyamine amide toxins) in spider andwasp venoms, and determined that some of the arylalkylamines present inthese venoms act as potent noncompetitive antagonists of glutamatereceptor-operated Ca²⁺ channels in the mammalian CNS. Although thesearylalkylamine compounds contain within their structure a polyaminemoiety, they are unlike other known simple polyamines in possessingextremely potent and specific effects on certain types ofreceptor-operated Ca²⁺ channels.

Using native arylalkylamines as lead structures, a number of analogswere synthesized and tested. Initial findings on arylalkylaminesisolated and purified from venom were confirmed utilizing syntheticarylalkylamines. These compounds are small molecules (mol. wt. <800)with demonstrated efficacy in in vivo models of stroke and epilepsy. TheNMDA receptor-ionophore complex was used as a model of receptor-operatedCa²⁺ channels. Selected arylalkylamines were shown to block NMDAreceptor-mediated responses by a novel mechanism. Moreover, the uniquebehavioral pharmacological profile of these compounds suggests that theyare unlikely to cause the PCP-like psychotomimetic activity andcognitive deficits that characterize other inhibitors of the NMDAreceptor. Finally, the arylalkylamines are unique amongst NMDA receptorantagonists in that they are able to antagonize certain subtypes ofcloned and expressed AMPA receptors, namely, those permeable to Ca²⁺.The arylalkylamines, therefore, are the only known class of compoundsable to antagonize glutamate receptor-mediated increases in cytosolicCa²⁺ regardless of the pharmacological definition of receptor subtype.Additionally, the arylalkylamines inhibit another receptor-operated Ca²⁺channel, the nicotinic cholinergic receptor. Given that excessive andprolonged increases in cytosolic Ca²⁺ have been implicated in theetiology of several neurological disorders and diseases, sucharylalkylamines are valuable small molecule leads for the development ofnovel therapeutics for various neurological disorders and diseases.

Applicant has determined that the selected arylalkylamines bind withhigh affinity at a novel site on the NMDA receptor-ionophore complexwhich has heretofore been unidentified, and that said arylalkylamines donot bind with high affinity at any of the known sites (glutamate bindingsite, glycine binding site, MK-801 binding site, Mg²⁺ binding site, Zn²⁺binding site, polyamine binding site, sigma binding site) on said NMDAreceptor-ionophore complex. This determination has allowed applicant todevelop methods and protocols by which useful compounds can beidentified which provide both therapeutically useful compounds and leadcompounds for the development of other therapeutically useful compounds.These compounds can be identified by screening for compounds that bindat this novel arylalkylamine binding site, and by determining whethersuch a compound has the required biological, pharmacological andphysiological properties.

The method includes the step of identifying a compound which binds tothe receptor-operated Ca²⁺ channel at that site bound by thearylalkylamine compounds referred to herein as Compound 1, Compound 2 orCompound 3, and having the structures shown below.

By “therapeutically useful compound” is meant a compound that ispotentially useful in the treatment of a disorder or disease state. Acompound uncovered by the screening method is characterized as havingpotential therapeutic utility in treatment because clinical tests havenot yet been conducted to determine actual therapeutic utility.

By “neurological disorder or disease” is meant a disorder or disease ofthe nervous system including, but not limited to, global and focalischemic and hemorrhagic stroke, head trauma, spinal cord injury, spinalcord ischemia, ischemia- or hypoxia-induced nerve cell damage,hypoxia-induced nerve cell damage as in cardiac arrest or neonataldistress, epilepsy, anxiety, neuropsychiatric or cognitive deficits dueto ischemia or hypoxia such as those that frequently occur as aconsequence of cardiac surgery under cardiopulmonary bypass, andneurodegenerative disease. Also meant by “neurological disorder ordisease” are those disease states and conditions in which aneuroprotectant, anticonvulsant, anxiolytic, analgesic, muscle relaxantand/or adjunct in general anesthesia may be indicated, useful,recommended or prescribed.

By “neurodegenerative disease” is meant diseases including, but notlimited to, Alzheimer's Disease, Huntington's Disease, Parkinson'sDisease, and amyotrophic lateral sclerosis (ALS).

By “neuroprotectant” is meant a compound capable of preventing theneuronal damage or death associated with a neurological disorder ordisease.

By “anticonvulsant” is meant a compound capable of reducing convulsionsproduced by conditions such as simple partial seizures, complex partialseizures, status epilepticus, and trauma-induced seizures such as occurfollowing head injury, including head surgery.

By “anxiolytic” is meant a compound capable of relieving the feelings ofapprehension, uncertainty and fear that are characteristic of anxiety.

By “analgesic” is meant a compound capable of relieving pain by alteringperception of nociceptive stimuli without producing anesthesia or lossof consciousness.

By “muscle relaxant” is meant a compound that reduces muscular tension.

By “adjunct in general anesthesia” is meant a compound useful inconjunction with anesthetic agents in producing the loss of ability toperceive pain associated with the loss of consciousness.

By “Potent” or “active” is meant that the compound has activity atreceptor-operated calcium channels, including NMDA receptors,Ca²⁺-permeable AMPA receptors, and nicotinic cholinergic receptors, withan IC₅₀ value less than 10 μM, more preferably less than 100 nM, andeven more preferably less than 1 nM.

By “selective,” is meant that the compound is potent atreceptor-operated calcium channels as defined above, but is less potentby greater than 10-fold, more preferably 50-fold, and even morepreferably 100-fold, at other neurotransmitter receptors,neurotransmitter receptor-operated ion channels, or voltage-dependention channels.

By “biochemical and electrophysiological assays of receptor-operatedcalcium channel function” is meant assays designed to detect bybiochemical or electrophysiological means the functional activity ofreceptor-operated calcium channels. Examples of such assays include, butare not limited to, the fura-2 fluorimetric assay for cytosolic calciumin cultured rat cerebellar granule cells (see Example 1 and Example 2),patch clamp electrophysiolocial assays (see Example 3 and Example 27),rat hippocampal slice synaptic transmission assays (see Example 5),radioligand binding assays (see Example 4, Example 24, Example 25, andExample 26), and in vitro neuroprotectant assays (see Example 6)

By “efficacy” is meant that a statistically significant level of thedesired activity is detectable with a chosen compound; by “significant”is meant a statistical significance at the p<0.05 level.

By “neuroprotectant activity” is meant efficacy in treatment ofneurological disorders or diseases including, but not limited to, globaland focal ischemic and hemorrhagic stroke, head trauma, spinal cordinjury, spinal cord ischemia, ischemia- or hypoxia-induced nerve celldamage, hypoxia-induced nerve cell damage as in cardiac arrest orneonatal distress, neuropsychiatric or cognitive deficits due toischemia or hypoxia such as those that frequently occur as a consequenceof cardiac surgery under cardiopulmonary bypass, and neurodegenerativediseases such as Alzheimer's Disease, Huntington's Disease, Parkinson'sDisease, and amyotrophic lateral sclerosis (ALS) (see Examples 7 and 8,below).

By “anticonvulsant activity” is meant efficacy in reducing convulsionsproduced by conditions such as simple partial seizures, complex partialseizures, status epilepticus, and trauma-induced seizures such as occurfollowing head injury, including head surgery (see Examples 9 and 10,below).

By “anxiolytic activity” is meant that a compound reduces the feelingsof apprehension, uncertainty and fear that are characteristic ofanxiety.

By “analgesic activity” is meant that a compound produces the absence ofpain in response to a stimulus that would normally be painful. Suchactivity would be useful in clinical conditions of acute and chronicpain including, but not limited to the following: preemptivepreoperative analgesia; peripheral neuropathies such as occur withdiabetes mellitus and multiple sclerosis; phantom limb pain; causalgia;neuralgias such as occur with herpes zoster; central pain such as thatseen with spinal cord lesions; hyperalgesia; and allodynia.

By “causalgia” is meant a painful disorder associated with injury ofperipheral nerves.

By “neuralgia” is meant pain in the distribution of a nerve or nerves.

By “central pain” is meant pain associated with a lesion of the centralnervous system.

By “hyperalgesia” is meant an increased response to a stimulus that isnormally painful.

By “allodynia” is meant pain due to a stimulus that does not normallyprovoke pain (see Examples 11 through 14, below).

By “induction of long-term potentiation in rat hippoc-ampal slices” ismeant the ability of tetanic electrical stimulation of afferent Schaffercollateral fibers to elicit long-term increases in the strength ofsynaptic transmission at the Schaffer collateral-CAl pyramidal cellpathway in rat hippocampal slices maintained in vitro (see Example 19).

By “therapeutic dose” is meant an amount of a compound that relieves tosome extent one or more symptoms of the disease or condition of thepatient. Additionally, by “therapeutic dose” is meant an amount thatreturns to normal, either partially or completely, physiological orbiochemical parameters associated with or causative of the disease orcondition. Generally, it is an amount between about 1 nmole and 1 μmoleof the compound, dependent on its EC₅₀ (IC₅₀ in the case of anantagonist) and on the age, size, and disease associated with thepatient.

By “impair cognition” is meant the ability to impair the acquisition ofmemory or the performance of a learned task (see Example 20). Also by“impair congnition” is meant the ability to interfere with normalrational thought processes and reasoning.

By “disrupt motor function” is meant the ability to significantly alterlocomotor activity (see Example 15) or elicit significant ataxia, lossof the righting reflex, sedation or muscle relaxation (see Example 16).

By “locomotor activity” is meant the ability to perform normalambulatory movements.

By “loss of the righting reflex” is meant the ability of an animal,typically a rodent, to right itself after being placed in a supineposition.

By “neuronal vacuolization” is meant the production of vacuoles inneurons of the cingulate cortex or retrosplenial cortex (see Example18).

By “cardiovascular activity” is meant the ability to elicit significantchanges in parameters including, but not limited to, mean arterial bloodpressure and heart rate (see Examples 21 and 22).

By “hyperexcitability” is meant an enhanced susceptibility to anexcitatory stimulus. Hyperexcitability is often manifested as asignificant increase in locomotor activity in rodents administered adrug (see Example 15).

By “sedation” is meant a calmative effect, or the allaying of activityand excitement. Sedation is often manifested as a significant decreasein locomotor activity in rodents administered a drug (see Example 15).

By “PCP-like abuse potential” is meant the potential of a drug to bewrongfully used, as in the recreational use of PCP (i.e., “angel dust”)by man. It is believed that PCP-like abuse potential can be predicted bythe ability of a drug to generalize to PCP in rodents trained todiscriminate PCP from saline (see Example 17.)

By “generalization to PCP” is meant that a compound is perceived asbeing PCP in rodents trained to discriminate PCP from saline (seeExample 17).

By “PCP-like psychotomimetic activity” is meant the ability of a drug toelicit in man a behavioral syndrome resembling acute psychosis,including visual hallucinations, paranoia, agitation, and confusion. Itis believed that PCP-like psychotomimetic activity can be predicted inrodents by the ability of a drug to produce PCP-like stereotypicbehaviors including ataxia, head weaving, hyperexcitability, andgeneralization to PCP in rodents trained to discriminate PCP from saline(see Example 15 Example 16, and Example 17).

By “ataxia” is meant a deficit in muscular coordination.

By “head weaving” is meant the stereotypic behavior elicited in rodentsby PCP in which the head is repeatedly moved slowly and broadly fromside to side.

By “pharmaceutical composition” is meant a therapeutically effectiveamount of a compound of the present invention in a pharmaceuticallyacceptable carrier, i.e., a formulation to which the compound can beadded to dissolve or otherwise facilitate administration of thecompound. Examples of pharmaceutically acceptable carriers includewater, saline, and physiologically buffered saline. Such apharmaceutical composition is provided in a suitable dose. Suchcompositions are generally those which are approved for use in treatmentof a specified disorder by the FDA or its equivalent in non-U.S.countries.

In a related aspect, the invention features a method for treating aneurological disease or disorder, comprising the step of administering apharmaceutical composition comprising a compound which binds to areceptor-operated calcium channel at the site bound by one of thearylalkylamines Compound 1, Compound 2 and Compound 3, said compoundbeing a potent and selective noncompetitive antagonist at such areceptor-operated calcium channel, and having one or more of thefollowing pharmacological and physiological properties:.efficacy in invitro biochemical and electrophysiological assay of receptor-operatedcalcium channel function, in vivo anticonvulsant activity, in vivoneuroprotectant activity, in vivo anxiolytic activity, and in vivoanalgesic activity; said compound also possessing one or more of thefollowing pharmacological effects: the compound does not interfere withthe induction of long-term potentiation in rat hippocampal slices, and,at a therapeutic dose, does not impair cognition, does not disrupt motorperformance, does not produce neuronal vacuolization, has minimalcardiovascular activity, does not produce sedation or hyperexcitability,has minimal PCP-like abuse potential, and has minimal PCP-likepsychotomimetic activity. By “minimal” is meant that any side effect ofthe drug is tolerated by an average individual, and thus that the drugcan be used for therapy of the target disease. Such side effects arewell known in the art and are routinely regarded by the FDA as minimalwhen it approves a drug for a target disease.

Treatment involves the steps of first identifying a patient that suffersfrom a neurological disease or disorder by standard clinical methodologyand then treating such a patient with a composition of the presentinvention.

In a further aspect, the invention features compounds useful fortreating a patient having a neurological disease or disorder whereinsaid compound is a polyamine-type compound or an analog thereof (i.e., apolyheteroatomic molecule) having the formula

wherein Ar is an appropriately substituted aromatic ring, ring system orother hydrophobic entity; Ar can be an aromatic (e.g., carbocyclic arylgroups such as phenyl and bicyclic carbocyclic aryl ring systems such asnaphthyl, 1,2,3,4-tetrahydronaphthyl, indanyl, and indenyl),heteroaromatic (e.g., indolyl, dihydroindolyl quinolinyl andisoquinolinyl, and their respective 1,2,3,4-tetrahydro- and2-oxo-derivatives), alicyclic (cycloaliphatic), or heteroalicyclic ringor ring system (mono-, bi-, or tricyclic), having 5- to 7-memberedring(s) optionally substituted with 1 to 5 substituents independentlyselected from lower alkyl of 1 to 5 carbon atoms, lower haloalkyl of 1to 5 carbon atoms substituted with 1 to 7 halogen atoms, lower alkoxy of1 to 5 carbon atoms, halogen, nitro, amino, lower alkylamino of 1 to 5carbon atoms, amido, lower alkylamido of 1 to 5 carbon atoms, cyano,hydroxyl, sulfhydryl, lower acyl of 2 to 4 carbon atoms, sulfonamido,lower alkylsulfonamido of 1 to 5 carbon atoms, lower alkylsulfoxide of 1to 5 carbon atoms, lower hydroxyalkyl of 1 to 5 carbon atoms, loweralkylketo of 1 to 5 carbon atoms, or lower thioalkyl of 1 to 5 carbonatoms,

each m is an integer from 0 to 3, inclusive,

each k is an integer from I to 10, inclusive,

each j is the same or different and is an integer from 1 to 12,inclusive,

each R¹ and R² independently is selected from the group consisting ofhydrogen, lower alkyl of 1 to 5 carbon atoms, lower alkylamino of 1 to 5carbon atoms, lower alkylamido of 1 to 5 carbon atoms, lower mono-, di-,or trifluoroalkyl of 1 to 5 carbon atoms, hydroxy, amidino, guanidino,or typical common amino acid side chain or with an associated carbonatom R¹ and R² taken together form a carbonyl, and

each Z is selected from the group consisting of nitrogen, oxygen,sulfur, amido, sulfonamido, and carbon.

Preferred aromatic headgroups include, but are not limited to, thefollowing:

Preferably the claims claiming a compound exclude known compounds whosechemical structures are enabled.

In further preferred embodiments, the compound is selected from thegroup of Compounds 4 through 18, where such compounds have the formulae:

Applicant has also determined (see Example 23 below) that simplifiedarylalkylamines (see below)are potent, noncompetitive antagonists of theNMDA receptor-ionophore complex. The simplified arylalkylamines aredistrict from the arylalkylamines exemplified by Compounds 4-18 asdescribed above. For example, such compounds bind to the site labeled by[³H]MK-801 at concentrations ranging approximately 1 to 400-fold higherthan those which antagonize NMDA receptor-mediated function. Suchsimplified arylalkylamines possess one or more of the followingadditional biological properties: significant neuroprotectant activity,significant anticonvulsant activity, significant analgesic activity, noPCP-like stereotypic behavior in rodents (hyperexcitability and headweaving) at effective neuroprotectant, anticonvulsant and analgesicdoses, no generalization to PCP in a PCP discrimination assay ateffective neuroprotectant, anticonvulsant and analgesic doses, noneuronal vacuolization at effective neuroprotectant, anticonvulsant andanalgesic doses, significantly less potent activity againstvoltage-sensitive calcium channels, and minimal hypotensive activity ateffective neuroprotectant, anticonvulsant and analgesic doses. Suchcompounds may, however, inhibit the induction of LTP in rat hippocampalslices and may produce motor impairment at neuroprotectant,anticonvulsant and analgesic doses.

One aspect of the invention features a method for treating a patienthaving a neurological disease or disorder, comprising administering acompound of Formula I:

wherein:

R¹ and R⁵ are independently selected from the group consisting ofphenyl, benzyl, and phenoxy (each of which is optionally substitutedwith alkyl, hydroxyalkyl, —OH, -0-alkyl, -0-acyl, —F, —Cl, —Br, —I,—CF₃, or —OCF₃) —H, alkyl, hydroxyalkyl, —OH, -0-alkyl, and 0-acyl;

R² and R⁶ are independently selected from the group consisting of —H,alkyl, and hydroxyalkyl; or R² and R⁶ together are imino; or R¹ and R²together are —(CH₂)_(n)— or —(CH₂)_(n)—N(R³)—(CH₂)_(n)—;

R³ is independently selected from the group consisting of —H, alkyl,2-hydroxyethyl and alkylphenyl;

n is an integer from 0 to 6, but at least one n must be greater than 0;

R⁴ is selected from the group consisting of thiofuran, pyridyl, phenyl,benzyl, phenoxy, and phenylthio (each of which is optionally substitutedwith alkyl, —F, —Cl, —Br, —I, —CF₃, —OH, —OCF₃, -0-alkyl, or -0-acyl),—H, alkyl and cycloalkyl;

X is independently selected from the group consisting of phenyl, benzyl,and phenoxy (each of which is optionally substituted with —F, —Cl, —Br,—I, —CF₃, alkyl, —OH, —OCF₃, -0-alkyl, or -0-acyl) —F, —Cl, —Br, —ICF₃,alkyl, —OH, —OCF₃, -0-alkyl, and 0-acyl;

m is independently an integer from 0 to 5;

Y is —NR³R³, except when R¹ and R² together are—(CH₂)_(n)—N(R³)—(CH₂)_(n)—, then Y is —H;

and pharmaceutically acceptable salts and complexes thereof, wherein thecompound is active at an NMDA receptor.

By “patient” is meant any animal that has a cell with an NMDA receptor.Preferably, the animal is a mammal. Most preferably, the animal is ahuman.

By “alkyl” is meant a branched or unbranched hydrocarbon chaincontaining between 1 and 6, preferably between 1 and 4, carbon atoms,such as, e.g., methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl,iso-butyl, tert-butyl, 2-methylpentyl, cyclopropylmethyl, allyl, andcyclobutylmethyl.

By “lower alkyl” is meant a branched or unbranched hydrocarbon chaincontaining between 1 and 4 carbon atoms, of which examples are listedherein.

By “hydroxyalkyl” is meant an alkyl group as defined above, substitutedwith a hydroxyl group.

By “alkylphenyl” is meant an alkyl group as defined above, substitutedwith a phenyl group.

By “acyl” is meant —C(O)R, where R is H or alkyl as defined above, suchas, e.g., formyl, acetyl, propionyl, or butyryl; or,

R is -0-alkyl such as in alkyl carbonates or R is N-alkyl such as inalkyl carbamates.

By “cycloalkyl” is meant a branched or unbranched cyclic hydrocarbonchain containing between 3 and 12 carbon atoms.

In preferred aspects of the invention, Y is selected from the groupconsisting of —NH₂ and —NH-methyl;

R⁴ is thiofuran, pyridyl, phenyl, benzyl, phenoxy, or phenylthio, eachof which is optionally substituted with —F, —Cl, —Br, —I, —CF_(3,)alkyl, —OH, —OCF₃, -0-alkyl, or -0-acyl;

X is independently selected from the group consisting of meta-fluaro,meta-chloro, ortho-O-lower alkyl, ortho-methyl, ortho-fluoro,ortho-chloro, meta-0lower alkyl, meta-methyl, ortho-OH, and meta-OH; andeither

R¹, R², R⁵, and R⁶ are —H;

or R is methyl, and R¹, R⁵, and R⁶ are —H;

or R¹ is methyl, and R², R⁵, and R⁶ are —H.

In other preferred aspects of the present invention,

R¹ and R⁵ are independently selected from the group consisting of —H,lower alkyl, hydroxyalkyl, —OH, -0-alkyl, and -0-acyl;.

R² and R⁶ are independently selected from the group consisting of —H,lower alkyl, and hydroxyalkyl;

or R¹ and R² together are —(CH₂)_(n)— or —(CH₂)_(n)—N(R³)—, and Y is H;

R³ is independently selected from the group consisting of —H and loweralkyl;

R⁴ is selected from the group consisting of thiofuran, pyridyl, phenyl,benzyl, phenoxy, and phenylthio (each of which is optionally substitutedwith lower alkyl, —F, —Cl, —Br, —I, —CF₃, —OH, —OCF₃, 0-alkyl, or-0-acyl), —H, lower alkyl, and cycloalkyl;

X is independently selected from the group consisting of —F, —Cl, —Br,—I, —CF₃, lower alkyl, —OH, and —OCF₃;

m is independently an integer from 0 to 5;

Y is —NHR³, or hydrogen when R¹ and R² together are —(CH₂)_(n)—N(R³)—,and pharmaceutically acceptable salts and complexes thereof, with theprovisos that

(a) when R¹ and R² together are —(CH₂)_(n)—N(R³)—, then R⁵, R⁶, and Yare hydrogens; and

(b) when R¹ and R² together are not (CH₂)_(n)—N(R³)—, then Y is —NHR³.

In one preferred aspect, the invention features a method for treating apatient having a neurological disease or disorder comprisingadministering a compound of Formula II:

wherein:

X is independently selected from the group consisting of —H, —Br, —Cl,—F, —I, —OCF₃, alky, —OH, —CCF₃, -0-alkyl, and -0-acyl;

R¹ is independently selected from the group consisting of —H. alkyl,hydroxyalkyl, —OH, -0-alkyl, and -0-acyl;

R² is independently selected from the group consisting of —H. alkyl, andhydroxyalkyl, or both R²s together are imino; R³ is independentlyselected from the group consisting of —H, alkyl, 2-hydroxyethyl, andalkylphenyl; and m is independently an integer from 0 to 5.

Thus, in this preferred aspect, the compounds include the compound ofFormula I, wherein:

X is independently selected from the group consisting of —F, —Cl, —Br,—I, —CF₃, alkyl, —OH, —OCF₃, -0-alkyl, and -0-acyl;

R¹ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, -0-alkyl, and -0-acyl;

R² and R⁶ are independently selected from the group consisting of —H,alkyl, and hydroxyalkyl, or R² and R⁶ together are imino;

R⁵ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, -O-alkyl, and -0-acyl;

Y is NR³R³; and

R⁴ is phenyl, optionally substituted with alkyl, —F, —Cl, —Br, —I, —CF₃,—OH, —OCF₃, -0-alkyl, or -0-acyl.

In another preferred aspect, the administered compound has the structureof Formula III:

wherein:

X is independently selected from the group consisting of —H, —Br, —Cl,—F, —I, —CF₃, alkyl, —OH, —OCF₃, -0-alkyl, and -0-acyl;

R¹ is independently selected from the group consisting of —H, alkyl,hydroxyalkyl, —OH, -0-alkyl, and -0-acyl;

R is independently selected from the group consisting of —H, alkyl, andhydroxyalkyl, or both R²s together are imino;

R³ is independently selected from the group consisting of —H, alkyl,2-hydroxyethyl, and alkylphenyl;

R⁴ is selected from the group consisting of thiofuran, pyridyl, phenyl,benzyl, phenoxy, and phenylthio, (each of which is optionallysubstituted with (X)m), alkyl, and cycloalkyl; and, m is independentlyan integer from 0 to 5.

Thus, in the-preferred aspect, the compounds include the compound ofFormula 1, wherein:

X is independently selected from the group consisting of —F, —Cl, —Br,—I, —OCF₃, alkyl, —OH, —OCF₃, -0-alkyl, and -0-acyl;

R¹ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, -0-alkyl, and -0-acyl;

R² and R⁶ are selected from the group consisting of —H, alkyl, andhydroxyalkyl, or R² and R⁶ together are imino;

R⁵ is independently selected from the group consisting of —H, alkyl,hydroxyalkyl, —OH, -0-alkyl, and -0-acyl; and

Y is NR³R³.

In another preferred aspect, the administered compound has the structureof Formulas IV and V.

wherein:

n is an integer from 1 to 6;

X is independently selected from the group consisting of —H, —Br, —Cl,—F, —I —CF3, alkyl, —OH, —OCF₃, -0-alkyl, and -0-acyl;

R³ is independently selected from the group consisting of —H, alkyl,2-hydroxyethyl, and alkylphenyl; and m is independently an integer from0 to 5.

Thus, in this preferred aspect, the administered compounds include thecompound of Formula I, wherein:

R³ is independently selected from the group consisting of —H, and alkyl;

R⁴ is phenyl, optionally substituted with alkyl, —F, —Cl, —Br, —I, —CF₃,—OH, —OCF₃, -0-alkyl, or -0-acyl; and

R1 and R2 together are —(CH₂)_(n),— or —(CH₂)_(n), —N(R³)—.

In another preferred aspect, the administered compound has the structureof Formulas VI and VII:

wherein:

n is an integer from 1 to 6;

X is independently selected from the group consisting of —H, —Br, —Cl,—F, —I, —CF, alkyl, —OH, —OCF₃, -0-alkyl, and -0-acyl;

R³ is independently selected from the group consisting of —H, alkyl,2-hydroxyethyl, and alkylphenyl;

R⁴ is selected from the group consisting of thiofuran, pyridyl, phenyl,benzyl, phenoxy, and phenylthio (each of which is optionally substitutedwith (X)m), alkyl, and cycloalkyl; and m is independently an integerfrom 0 to 5.

Thus, in this preferred aspect, the administered compounds include thecompound of Formula I, wherein:

X is independently selected from the group consisting of —F, —Cl, —Br,—I, CF3, alkyl, —OH, —OCF3, -0-alkyl, and -0-acyl-; and

R₁ and R₂ together are —(CH₂)_(n)— or —(CH₂)_(n)—N(R³)—.

More preferred aspects are those embodiments in which:

X is independently selected from the group consisting of meta-fluoro,meta-chloro, ortho-O-lower alkyl, ortho-methyl, ortho-fluoro,ortho-chloro, meta-O-lower alkyl, meta-methyl, ortho-OH, and meta-OH;

NH³ is selected from the group consisting of NH, N-methyl, and N-ethyl;

NR³R³ is selected from the group consisting of NH₂, NH-methyl, andNH-ethyl;

R¹ is selected from the group consisting of —H and methyl;

R² is selected from-the group consisting of —H and methyl; and

R⁴ is selected from the group consisting of phenyl, benzyl, and phenoxy,each of which is optionally substituted with alkyl, —F, —Cl, —Br, —F,—CF₃, —OH, —OCF₃, -0-alkyl, or -0-acyl.

Especially preferred aspects are those embodiments in which:

X is meta-fluoro;

each R¹ and R² is —H;

NR³ is selected from the group consisting of NH and N-methyl;

NR³R³ is selected from the group consisting of NH₂ and NH-methyl; and

R⁴ is selected from the group consisting of phenyl, benzyl, and phenoxy,each of which is optionally substituted with alkyl, —F, —Cl, —Br, —I,—CF₃, —OH, —OCF₃, -0-alkyl, or -0-acyl.

In a further aspect, the invention features a method for treating apatient having a neurological disease or disorder comprisingadministering the compounds of Formula VIII:

wherein:

Z is selected from the group consisting of —CH₂CH₂—, —CH₂CH(CH₃)—,—CH═CH—, -0-CH₂—, S—CH₂—, -0, and —S—;

X¹ and X² are independently selected from the group consisting of —F,—Cl, —CH₃, —OH, and lower 0-alkyl in the 1-, 3-, 7-, or 9-substituentpositions;

m is independently an integer from 0 to 2;

—NHR is selected from the group consisting of —NH₂, —NHCH₃, and —NHC₂H₅;

R¹ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, —O-alkyl, and —O-acyl, and

R² is selected from the group consisting of —H, alkyl, hydroxyalkyl, andpharmaceutically acceptable salts and complexes thereof, wherein thecompound is active at an NMDA receptor.

Especially preferred aspects are those embodiments in which:

Z is —CH₂CH₂—;

X¹ or X² is —F, or both X¹ and X² are —F;

either R¹ or R² is methyl or both R¹ and R² are —H; and

-   -   —NHR is selected from the group consisting of —NH₂ or —NHCH₃.

In other preferred embodiments, the invention features—a method fortreating a patient having a neurological disease or disorder comprisingadministering the compounds of Formula IX:

wherein:.

W is selected from the group consisting of —CH₂—, —O—, and —S—;

X¹ and X² are independently selected from the group consisting of —F,—Cl, —CH₃, —OH, and lower 0-alkyl;

m is independently an integer from 0 to 2;

—HR is selected from the group consisting of —NH₂, —NHCH₃, and —NHC₂H₅;

R¹ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, -0-alkyl, and -0-acyl; and

R² is selected from the group consisting of —H, alkyl, hydroxyalkyl, andpharmaceutically acceptable salts and complexes thereof, wherein thecompound is active at an NMDA receptor.

In preferred aspects, the administered compound is selected from thegroup consisting of Compound 128, 129, 130, 131, 132, 200, 201, 202,203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, and 215.

In preferred embodiments, the methods of treatment includeadministration of a compound selected from Compounds 19 through 215, orpharmaceutically acceptable salts and complexes thereof. Preferably, thecompound has an IC₅₀≦10 μM at an NMDA receptor, more preferably ≦2.5 μM,and most preferably ≦0.5 μM at an NMDA receptor.

In further preferred embodiments, the methods of treatment includeadministration of a compound selected from the group consisting ofCompound 19, 20, 21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,75, 76, 77, 78, 79, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 100, 101, 102, 103, 105, 106, 107, 108, 109, 111,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138 (potentialprodrug), 139, 141, 142, 143, 144, 145, 146, 147, 148, 149, and 150, andpharmaceutically acceptable salts and complexes thereof. These compoundshave an IC₅₀≦10 μm at an NMDA receptor.

In more preferred embodiments, the methods of treatment includeadministration of a compound selected from the group consisting ofCompound 19, 20, 21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 70, 75, 76, 81, 82, 83,85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 100, 101, 102, 103,105, 106, 108, 109, 111, 115, 118, 119, 120, 121, 122, 125, 126, 127,128, 129, 130, 131, 132, 133, 135, 136, 137, 138 (potential prodrug),139, 142, 144, 145, 146, 147, 148, 149, and 150, and pharmaceuticallyacceptable salts and complexes thereof. These compounds have an IC₅₀≦2.5μM at an NMDA receptor.

In other embodiments, the compound is selected from the group consistingof Compound 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 76,82, 83, 88, 89, 90, 92, 93, 94, 95, 96, 101, 102, 103, 105, 109, 111,115, 118, 119, 120, 121, 122, 125, 126, 127, 129, 130, 131, 135, 136,137, 138, 139, 142, 144, 145, 148, 149, and 150, and pharmaceuticallyacceptable salts and complexes thereof.

In particularly preferred embodiments, the methods of treatment includeadministration of a compound selected from the group consisting ofCompound 19, 20, 21, 22, 23, 24, 25, 27, 28, 30, 31, 32, 33, 38, 39, 43,44, 46, 47, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 69, 82, 83, 89, 90, 91, 93, 94, 95, 96, 97, 103, 111, 118, 119,120, 122, 126, 135, 136, 137, 138 (potential prodrug), 142, 144, 145,147, 148, 149, and 150, and pharmaceutically acceptable salts andcomplexes thereof. These compounds have an IC₅₀≦0.5 μM at an NMDAreceptor.

In more preferred embodiments, the methods of treatment includeadministration of a compound selected from the group consisting ofCompound 20, 24, 25, 33, 50, 60, 66, 69, 103, 111, 118, 119, 120, 122,136, 137, 138 (potential prodrug), 142, 144, 145, 148, 149, and 150, andpharmaceutically acceptable salts and complexes thereof.

In particularly preferred embodiments, the methods of treatment includeadministration of a compound selected from the group consisting ofCompound 20, 33, 50, 60, 119, and 144, and pharmaceutically acceptablesalts and complexes thereof.

In other particularly preferred embodiments, the methods of treatmentinclude administration of a compound selected from the group consistingof Compound 33, 50, 60, 119, and 144, and pharmaceutically acceptablesalts and complexes thereof.

In preferred aspects, the invention provides a method for treating apatient having a neurological disease or disorder, comprisingadministering a compound which is selected from the group consisting ofCompound 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167,168, 169, 170, 171, 181, 182, 183, 184, 185, 186, 187, andpharmaceutically acceptable salts and complexes thereof. These compoundshave an IC₅₀≦10 μM at an NMDA receptor.

In further preferred aspects, the invention provides a method fortreating a patient having a neurological disease or disorder, comprisingadministering a compound which is selected from the group consisting ofCompound 157, 158, 159, 163, 164, 166, 167, 168, 169, 170, 171, 181,185, 186, and pharmaceutically acceptable salts and complexes thereof.These compounds have an IC₅₀≦10 μM at an NMDA receptor.

In other more preferred aspects, the invention provides a method fortreating a patient having a neurological disease or disorder, comprisingadministering a compound which is selected from the group consisting ofCompound 156, 157, 158, 159, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 181, 183, 184, 185, 186, 187, and pharmaceuticallyacceptable salts and complexes thereof. These compounds have anIC_(50≦2.5) μM at an NMDA receptor.

In further preferred aspects, the invention provides a method fortreating a patient having a neurological disease or disorder, comprisingadministering a compound which is selected from the group consisting ofCompound 157, 158, 159, 163, 164, 166, 167, 168, 169, 170, 171, 181,185, 186, and pharmaceutically acceptable salts and complexes thereof.These compounds have an IC₅₀≦2.5 μM at an NMDA receptor.

In other particularly preferred aspects, the invention provides a methodfor treating a patient having a neurological disease or disorder,comprising administering a compound which is selected from the groupconsisting of Compound 156, 157, 158, 159, 161, 163, 164, 165, 167, 168,169, 170, 171, 181, 186 and pharmaceutically acceptable salts andcomplexes thereof. These compounds have an IC₅₀≦0.5 μM at an NMDAreceptor.

In further preferred aspects, the invention provides a method fortreating a patient having a neurological disease or disorder, comprisingadministering a compound which is selected from the group consisting ofCompound 157, 158, 159, 163, 164, 167, 168, 169, 170, 171, 181, 186 andpharmaceutically acceptable salts and complexes thereof. These compoundshave an IC₅₀≦0.5 μM at an NMDA receptor.

In other preferred aspects, the invention provides a method for treatinga patient having a neurological disease or disorder comprising ofadministering a compound selected from the group consisting of Compounds151-215, and pharmaceutically acceptable salts and complexes thereof.

In more preferred aspects, the invention provides a method for treatinga patient having a neurological disease or disorder comprisingadministering a compound selected from the group consisting of Compound151, 152, 153, 154, 155, 157, 158, 159, 163, 164, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 181, 185, 186, 188, 189, 190, 191,192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205,206, 207, 208, 209, 210, 211, 212, 213, 214, 215 and pharmaceuticallyacceptable salts and complexes thereof.

The present invention provides simplified arylalkylamines comprising thecompounds of Formulas I-IX and all preferred aspects of Formulas I-IX asset out above.

Examples of such simplified arylalkylamines include, but are not limitedto, Compounds 19-215, whose structures are provided below. Preferably,the compound has an IC₅₀≦10 μM at an NMDA receptor. More preferably, thecompound has an IC₅₀≦5 μM, more preferably ≦2.5 μM, and most preferably≦0.5 μM at an NMDA receptor.

In preferred embodiments, the compound is selected from the groupconsisting of Compound 21, 22, 23, 24, 25, 26, 27, 28, 29, 33, 34, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 76, 78, 79, 82, 83, 84,88, 89, 90, 92, 93, 94, 95, 96, 98, 101, 102, 103, 105, 107, 108, 109,111, 115, 116, 118, 119, 120, 121, 122, 124, 125, 126, 127, 129, 130,131, 134, 135, 136, 137, 138 (potential prodrug), 139, 141, 142, 143,144, 145, 148, 149, and 150. These compounds have an IC₅₀≦10 μM at anNMDA receptor.

In other embodiments, the compound is selected from the group consistingof Compound 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 76,82, 83, 88, 89, 90, 92, 93, 94, 95, 96, 101, 102, 103, 105, 109, 111,115, 118, 119, 120, 121, 122, 125, 126, 127, 129, 130, 131, 135, 136,137, 138, 139, 142, 144, 145, 148, 149, and 150.

In more preferred embodiments, the compound is selected from the groupconsisting of Compound 21, 22, 23, 24, 25, 27, 28, 29, 33, 34, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69; 76, 82, 83, 88, 89, 90, 92,93, 94, 95, 96, 101, 102, 103, 105, 108, 109, 111, 115, 118, 119, 120,121, 122, 125, 126, 127, 129, 130, 131, 135, 136, 137, 138 (potentialprodrug), 139, 142, 144, 145, 148, 149, and 150. These compounds have anIC₅₀≦2.5 μM at an NMDA receptor.

In particularly preferred embodiments, the compound is selected from thegroup consisting of Compound 21, 22, 23, 24, 25, 27, 28, 33, 38, 39, 43,44, 46, 47, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 69, 82, 83, 89, 90, 93, 94, 95, 96, 103, 111, 118, 119, 120,122, 126, 135, 136, 137, 138. (potential prodrug), 142, 144, 145, 148,149, and 150. These compounds have an IC₅₀≦0.5 μM at an NMDA receptor.

In preferred embodiments, the compound is selected from the groupconsisting of Compound 24, 25, 33, 50, 60, 66, 69, 103, 111, 118, 119,120, 122, 136, 137, 138, 142, 144, 145, 148, 149, and 150.

In particularly preferred embodiments, the compound is selected from thegroup consisting of Compound 20, 33, 50, 60, 119, and 144.

In more particularly preferred embodiments, the compound is selectedfrom the group consisting of Compound 33, 50, 60, 119, and 144.

In other preferred aspects, the compound is selected from the groupconsisting of Compound 151, 152, 153, 154, 155, 157, 158, 159, 163, 164,166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 181, 185, 186,188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201,202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215 andpharmaceutically acceptable salts and complexes thereof.

In other preferred aspects, the compound is selected from the groupconsisting of Compound 157, 158, 159, 163, 164, 166, 167, 168, 169, 170,171, 181, 185, 186, and pharmaceutically acceptable salts and complexesthereof. These compounds have an IC₅₀≦10 μM at an NMDA receptor.

In more preferred aspects, the compound is selected from the groupconsisting of Compound 157, 158, 159, 163, 164, 167, 168, 169, 170, 171,131, 185, 186, and pharmaceutically acceptable salts and complexesthereof. These compounds have an IC₅₀≦2.5 μM at an NMDA receptor.

In most preferred aspects, the compound is selected from the groupconsisting of Compound 157, 158, 159, 163, 164, 167, 168, 169, 170, 171,181, 186, and pharmaceutically acceptable salts and complexes thereof.These compounds have an IC₅₀≦0.5 μM at an NMDA receptor.

Excluded from the composition of matter aspect of the present inventionare known compounds whose chemical structures are covered by the genericformulae presented above.

Also provided in an aspect of the invention are pharmaceuticalcompositions useful for treating a patient having a neurological diseaseor disorder. The pharmaceutical compositions are provided in apharmaceutically acceptable carrier and appropriate dose. Thepharmaceutical compositions may be in the form of pharmaceuticallyacceptable salts and complexes, as is known to those skilled in the art.

The pharmaceutical compositions comprise the compounds of Formulas I-IXand all preferred aspects of Formulas I-IX as set out above.

Preferred pharmaceutical compositions comprise Compounds 19-215.Preferably, the compound has an IC₅₀≦10 μM at an NMDA receptor. Morepreferably the compound has an IC₅₀≦5 μM, more preferably ≦2.5 μM, andmost preferably ≦0.5 μM at an NMDA receptor.

In further preferred embodiments, the pharmaceutical compositioncomprises a compound selected from the group consisting of Compound 20,21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 75, 76, 77, 78,79, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 100, 101, 102, 103, 105, 106, 107, 108, 109, 111, 114, 115, 116,117, 118, 119, 120, 121, 122, 213, 124, 125, 126, 127, 128, 129, 130,131, 132, 133, 134, 135, 136, 137, 138, (potential-prodrug), 139, 141,142, 143, 144, 145, 146, 147, 148, 149, and 150. These compounds have anIC₅₀≦10 μM at an NMDA receptor.

Preferably, the compound is selected from the group consisting of 21,22, 23, 24, 25, 26, 27, 28, 29, 33, 34, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 69, 76, 78, 79, 82, 83, 84, 88, 89, 90, 92, 93, 94, 95,96, 98, 101, 102, 103, 105, 107, 108, 109, 111, 115, 116, 118, 119, 120,121, 122, 124, 125, 126, 127, 129, 130, 131, 134, 135, 136, 137, 138(potential prodrug), 139, 141, 142, 143, 144, 145, 148, 149, and 150.

In other embodiments, the compound is selected from the group consistingof 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 76, 82, 83,88, 89, 90, 92, 93, 94, 95, 96, 101, 102, 103, 105, 109, i11, 115, 118,119, 120, 121, 122, 125, 126, 127, 129, 130, 131, 135, 136, 137, 138,139, 142, 144, 145, 148, 149, and 150.

In more preferred embodiments, the pharmaceutical composition comprisesa compound selected from the group consisting of Compound 20, 21, 22,23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 69, 70, 75, 76, 81, 82, 83, 85, 86, 871 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 100, 101, 102, 103, 105, 106, 108, 109, 111,115, 118, 119, 120, 121, 122, 125, 126, 127, 128, 129, 130, 131, 132,133, 135, 136, 137, 138 (potential prodrug), 139, 142, 144, 145, 146,148, 149, and 150. These compounds have an IC₅₀≦2.5 μM at an NMDAreceptor.

Preferably, the compound is selected from the group consisting of 21,22, 23, 24, 25, 27, 28, 29, 33, 34, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 53, 59, 60, 61, 62, 63,64, 65, 66, 69, 76, 82, 83, 88, 89, 90, 92, 93, 94, 95, 96, 101, 102,103, 105, 108, 109, 111, 115, 118, 119, 120, 121, 122, 125, 126, 127,129, 130, 131, 135, 136, 137, 138 (potential prodrug), 139, 142, 144,145, 148, 149, and 150.

In particularly preferred embodiments, the pharmaceutical compositioncomprises a compound is selected from the group consisting of Compound20, 21, 22, 23, 24, 25, 27, 28, 30, 31@, 32, 33, 38, 39, 43, 44, 46, 47,49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69,82, 83, 89, 90, 911 93, 94, 95, 96, 97, 103, 111, 118, 119, 120, 122,126, 135, 136, 137, 138 (potential prodrug), 142, 144, 145, 148, 149,and 150. These compounds leave an IC₅₀≦0.5, μM at an NMDA receptor.

Preferably, the compound is selected from the group consisting of 21,22, 23, 24, 25, 27, 28, 33, 38, 39, 43, 44, 46, 47, 49, 50, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 69, 82, 83, 89, 90, 93,94, 95, 96, 103, 111, 118, 119, 120, 122, 126, 135, 136, 137, 138(potential prodrug), 142, 144, 145, 148, 149, and 150.

In more preferred embodiments, the pharmaceutical composition comprisesa compound selected from the group consisting of Compound 20, 24, 25,33, 50, 60, 66, 69, 103, 111, 118, 119, 120, 122, 136, 137, 138, 142,144, 145, 148, 149, and 150.

Preferably, the compound is selected from the group consisting ofCompound 24, 25, 33, 50, 60, 66, 69, 103, 111, 118, 119, 120, 122, 136,137, 138, 142, 144, 145, 148, 149, and 150.

In most particularly preferred embodiments, the pharmaceuticalcomposition comprises a compound selected from the group consisting ofCompound 20, 33, 50, 60, 119, and 144.

Preferably, the compound is selected from the group consisting of 33,50, 60, 119, and 144.

In other preferred aspects, the pharmaceutical composition comprises acompound selected from the group consisting of compound 151, 152, 153,154, 155, 157, 158, 159, 163, 164, 166, 167, 168, 169, 170, 171, 172,173, 174, 175, 176, 181, 135, 186, 188, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215 and pharmaceutically acceptable saltsand complexes thereof, and a pharmaceutically acceptable carrier.

In other preferred aspects the pharmaceutical composition comprises acompound which is selected from the group consisting of Compound 157,158, 159, 163, 164, 166, 167, 168, 169, 170, 171, 181, 185, 186, andpharmaceutically acceptable salts and complexes thereof, and apharmaceutically acceptable carrier. These compounds have an IC₅₀≦10 μMat an NMDA receptor.

In more preferred aspects, the pharmaceutical composition comprises acompound which is selected from the group consisting of Compound 157,158, 159, 163, 164, 166, 167, 168, 169, 170, 171, 181, 185, 186, andpharmaceutically acceptable salts and complexes thereof, and apharmaceutically acceptable carrier. These compounds have an IC₅₀≦2.5 μMat an NMDA receptor.

In most preferred aspects, the pharmaceutical composition comprises acompound which is selected from the group consisting of Compound 157,158, 159, 163, 164, 167, 168, 169, 170, 171, 181, 186, andpharmaceutically acceptable salts and complexes thereof, and apharmaceutically acceptable carrier. These compounds have an IC₅₀≦0.5 μMat an NMDA receptor.

Structural modifications can be made to compounds such as 20 or 60 whichdo not add materially to the structure-activity relationships (SAR)illustrated here. For example, successful bioisosteric replacement orsubstitution of optionally substituted phenyl groups, such as thoseoccurring in Compounds 20 or 60, can be accomplished with otherlipophilic or semi-polar aromatic (e.g., naphthyl, naphthoxy, benzyl,phenoxy, phenylthio), aliphatic (alkyl, e.g., isopropyl), cycloaliphatic(cycloalkyl, e.g., cyclohexyl), heterocyclic [e.g., pyridyl, furanyl,thiofuranyl (thiophenyl)], or other functional groups or systems, as iswell known in the art, will afford clinically useful compounds(structural homologs, analogs, and/or congeners) with similarbiopharmaceutical properties and activity at the NMDA receptor (e.g.,cf. Compounds 37, 75, 79, 83, 89, 119-122, 125, 126, 128, 130, 132, 137,144, and 145). For example, such replacements or substitutions have beenused to great effect in the development of SAR among other groups ofhighly clinically and commercially successful synthetic pharmaceuticalagents such as the classical H₁-antihistamines, anticholinergics(antimuscarinics; e.g., anti-Parkinsonians), antidepressants (includingtricyclic compounds), and opioid analgesics [See, Foye et al. (Eds.),Principles of Medicinal Chemistry, 4th ed., Lea and Febiger/Williams andWilkins, Philadelphia, Pa., 1995, pp. 233, 265, 281-282, 340-341,418-427, and 430; Prous, J. R., The Year's Drug News, TherapeuticTargets—1995 Edition, Prous Science Publishers, Barcelona, Spain, 1995,pp. 13, 55-56, 58-59, 74, 89, 144-145, 152, 296297, and 317]. Similarly,bioisosteric replacement or substitution of the methylene or methinegroups in the propyl backbone of compounds such as 20 or 60 with, e.g.,oxygen, sulfur, or nitrogen, will afford clinically useful NMDAreceptor-active compounds with similarly useful biopharmaceuticalproperties, such as Compound-88 (a modified “classicalH₁-antihistamine-type” structure), which can be further optimized foractivity at the NMDA receptor by preparing, e.g., the correspondingcompounds) containing, e.g., (bis)(3-fluorophenyl) group(s), as taughtby the present invention. The propyl backbone of compounds such as 20and 60 may also be modified successfully by the incorporation of ringsystems (as in Compounds 102 and 111) and/or unsaturation (e.g., adouble bond, as in Compounds 81, 106, 109, and 139) to afford furtherclinically useful NMDA receptor-active compounds of the presentinvention (cf. compounds cited above).

In a related aspect, the invention features a method for making atherapeutic agent comprising the steps of screening for said agent bydetermining whether said agent is active on a receptor-operated calciumchannel, and synthesizing said therapeutic agent in an amount sufficientto provide said agent in a therapeutically effective amount to apatient. Said screening may be performed by methods known to those ofordinary skill in the art, and may, for example be performed by themethods set out herein. Those skilled in the art are also familiar withmethods used to synthesize therapeutic agents in amounts sufficient tobe provided in a therapeutically effective amount.

In a preferred aspect, said receptor-operated calcium channel is an NMDAreceptor. In a more preferred aspect, said method further comprises thestep of adding a pharmaceutically acceptable carrier to said agent. In afurther preferred aspect said therapeutic agent comprises a compound ofFormula I, as set out herein. In a further preferred aspect saidtherapeutic agent comprises a compound of Formula II, III, IV, V, VI,VII, VIII, or IX, as set out herein. In particularly preferred aspects,said therapeutic agent comprises a compound having a structure selectedfrom the group consisting of Formulas I-IX, and all preferred aspects ofsaid formulas as set out herein. In further preferred aspects, saidtherapeutic agent is selected from the group consisting of Compounds19-215. In a particularly preferred aspect, said therapeutic agent isprovided to a patient having a neurological disease or disorder. In arelated aspect, said screening comprises the step of identifying acompound which binds to said receptor-operated calcium channel at a sitebound by one of the arylalkylamines Compound 1, Compound 2, and Compound3.

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments thereof, and from theclaims.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following is a detailed description of the methods and tests bywhich therapeutically useful compounds can be identified and utilizedfor the treatment of neurological disorders and diseases. The tests areexemplified by use of Compound 1, Compound 2 or Compound 3, but othercompounds which have similar biological activity in these assays canalso be used (as discovered) to improve on the tests. Lead compoundssuch as Compound 1, Compound 2 or Compound 3 can be used for molecularmodeling using standard procedures, or existing or novel compounds innatural product libraries can be screened by the methods describedbelow.

One key method is the means by which compounds can be quickly screenedwith standard radioligand binding techniques (a radiolabeledarylalkylamine binding assay) to identify those which bind at the samesite on receptor-operated Ca²⁺ channels as Compound 1, Compound 2 orCompound 3. Data from such radioligand binding studies will also confirmthat said compounds do not inhibit [³HI arylalkylamine binding via anaction at the known sites on receptor-operated Ca²⁺ channels (such asthe glutamate binding site, glycine binding site, MK-801 binding site,Zn²⁺ binding site, Mg²⁺ binding site, sigma binding site, or polyaminebinding site on the NMDA receptor-ionophore complex). This screeningtest allows vast numbers of potentially useful compounds to beidentified and screened for activity in the other assays. Those skilledin the art will recognize that other rapid assays for detection ofbinding to the arylalkylamine site on receptor-operated Ca²⁺ channelscan be devised and used in this invention.

Additional testing utilizes electrophysiological (patch clamp)methodology to extend the results obtained with the above-mentionedradioligand binding assay. Such results will confirm that compoundsbinding to the arylalkylamine site are functional, noncompetitiveantagonists of receptor-operated Ca²⁺ channels with the followingproperties in common with the arylalkylamines themselves: open-channelblock manifested as use-dependent block, and voltage-dependent onset andreversal from block. Such results will also confirm that said compoundsdo not have their primary activity at the previously described sites onreceptor-operated Ca²⁺ channels (such as the glutamate binding site,glycine binding site, MK-801 binding site, Zn²⁺ binding site, Mg²⁺binding site, sigma binding site, or polyamin binding site on the NMDAreceptor-ionophore complex).

In addition, recombinant DNA technology can be used to make such testingeven more rapid. For example, using standard procedures, the gene(s)encoding the novel arylalkylamine binding site (i.e., receptor) can beidentified and cloned. This can be accomplished in one of several ways.For example, an arylalkylamine affinity column can be prepared, andsolubilized membranes from cells or tissues containing thearylalkylamine receptor passed over the column. The receptor moleculesbind to the column and are thus isolated. Partial amino acid sequenceinformation is then obtained which allows for the isolation of the geneencoding the receptor. Alternatively, CDNA expression libraries areprepared and subtractions of the library are tested for their ability toimpart arylalkylamine receptors on cells which do not normally expresssuch receptors (e.g., CHO cells, mouse L cells, HEK 293 cells, orXenopus oocytes). In this way, the library fraction containing the cloneencoding the receptor is identified. Sequential subfractionation ofactive library fractions and assay eventually results in a single cloneencoding the arylalkylamine receptor. Similarly, hybrid-arrest orhybrid-depletion cloning can be used. Xenopus oocytes are injected withMRNA from a appropriate tissue or cell source (e.g., human braintissue). Expression of the arylalkylamine receptor is detected as, forexample, an NMDA- or glutamate-stimulated influx of calcium which can beblocked by Compound 1, Compound 2 or Compound 3. CDNA clones are testedfor their ability to block expression of this receptor when CDNA or CRNAare hybridized to the mRNA of choice, prior to injection into Xenopusoocytes. The clone responsible for this effect is then isolated by theprocess described above once the receptor gene is isolated, standardtechniques are used to identify the polypeptide or portion(s) thereofwhich is(are) sufficient for binding arylalkylamines (the arylalkylaminebinding domain[s]). Further, using standard procedures, the entirereceptor or arylalkylamine binding domain(s) can be expressed byrecombinant technology. Said receptor or binding domain(s) can beisolated and used as a biochemical reagent such that, rather than usinga competitive assay exemplified below, a simple direct binding assay canbe used. That is, a screen is set up for compounds which bind at thenovel arylalkylamine receptor. In this way large numbers of compoundscan be simultaneously screened, e.g., by passage through a columncontaining the novel arylalkylamine receptor or arylalkylamine bindingdomain, and analysis performed on compounds which bind to the column.

Additional testing utilizes the combination of molecular biologicaltechniques (expression of cloned NMDA, AMPA or nicotinic cholinergicreceptors) and patch clamp electrophysiological techniques.Specifically, arylalkyl-amine analogs can be rapidly screened forpotency at cloned and expressed subunits of the above-mentionedreceptor-ionophore complexes. Site-directed mutagenesis can be utilizedin an effort to identify which amino acid-residues may be important indetermining arylalkylamine potency.

Assays for Potent and Selective Antagonists of Receptor-Operated CalciumChannels in the Mammalian CNS

Desired properties of a drug include: high affinity and selectivity forreceptor-operated Ca²⁺ channels, such as those present in NMDA, AMPA andnicotinic cholinergic receptor-ionophore complexes (compared toresponses mediated—via other neurotransmitter receptors,neurotransmitter receptor-operated ion channels, or voltage-dependention channels) and a noncompetitive antagonism of said receptor-operatedCa²⁺ channels.

The NMDA receptor-ionophore complex is utilized as an example of areceptor-operated Ca 2, channel. Activation of the NMDA receptor opens acation-selective channel that allows the influx of extracellular Ca²⁺and Na⁺, resulting in increases in [Ca²⁺] and depolarization of the cellmembrane. Measurements of [Ca²⁺], were used as primary assays fordetecting the activity of arylalkylamine compounds on NMDA receptors.Purified arylalkylamines, synthetic aryl-alkylamines, and syntheticanalogs of arylalkylamines were examined for activity in in vitro assayscapable of measuring glutamate receptor activity. Selected for detailedstudy were the arylalkylamines present in the venom of various spiderspecies. The arylalkylamines present in these venoms are structurallydistinct but have the basic structure of the class-represented byCompounds 1 through 3. Other more simplified synthetic analogs generallyconsist of suitably substituted aromatic chromophobic groups attached toan alkyl(poly)amine moiety (see Compounds 19 through 215 below).

A primary assay that provides a functional index of glutamate receptoractivity and that allows high-throughput screening was developed.Primary cultures of rat cerebellar granule cells loaded with thefluorimetric indicator fura-2 were used to-measure changes in [Ca²⁺]_(I)elicited-by NMDA and its coagonist glycine. This assay provides anextremely sensitive and precise index of NMDA receptor activity.Increases in [Ca²⁺]_(I) evoked by NMDA are dependent on the presence ofglycine, and are blocked by extracellular Mg²⁺ or antagonists acting atthe glutamate, glycine, or MK-801 binding sites. Increases in [Ca²⁺]elicited by NMDA/glycine are readily distinguished from those resultingfrom depolarization by their refractoriness to inhibition by blockers ofvoltage-sensitive Ca²⁺, channels. The fidelity with which measurementsof Ca²⁺], corroborate results obtained by electrophysiological andligand-binding studies suggests that such measurements mirror closelyactivation of the NMDA receptor-ionophore complex.

EXAMPLE 1 Potent Noncompetitive Inhibition of NMDA Receptor Function

Preferential inhibitory effects of arylalkylamines on NMDAreceptor-mediated increases in [Ca²⁺]_(I) in cultured rat cerebellargranule cells were measured. Increases in [Ca²⁺]_(I) were elicited bythe addition of NMDA/glycine (50 μM/1 μM) in the presence or absence ofdifferent concentrations of each test compound. The IC₅₀ values werederived for each test compound using from 2 to 8 separate experimentsper test compound, and the standard error level was less than 10% of themean value for each compound.

All of the arylalkylamines tested blocked increases in [Ca²⁺]_(i) incerebellar-granule cells elicited by NMDA/glycine. Certainarylalkylamines similar in structure to Compound 1 or Compound 2 werenearly as potent as MK-801 (IC₅₀=34 nM) which is the most potentcompound in the literature known to preferentially block NMDA receptors.Compound 3 had an IC₅₀=2 nM, that is, 17-fold more potent than MK-801.Many of the arylalkylamines tested were more potent than competitiveantagonists such as APS (IC₅₀=15 μM). The inhibitory effects of thearylalkylamines were not overcome by increasing the concentrations ofNMDA or glycine. That is, no change was observed in the EC₅₀ for eitherNMDA or glycine. The arylalkylamines are thus noncompetitive antagonistsat the NMDA receptor-idnophore complex, and act neither at the glutamatenor the glycine binding sites.

EXAMPLE 2 Activity Against Kainate and AMPA Receptor Function

Measurements of [Ca²⁺]_(I) in cerebellar granule cells can also be usedto monitor activation of the native kainate or AMPA receptors present inthis tissue. Although the increases in [Ca²⁺]_(I) evoked by theseagonists are of a lesser magnitude than those evoked by NMDA/glycine,such responses are robust and can be used to precisely assess thespecificity of action of arylalkylamines on pharmacologically definedglutamate receptor subtypes. Comparative measurements of [Ca²⁺]_(I)revealed a clear distinction in the receptor selectivity of thearylalkylamines. Some, like JSTX-3 (Joro Spider toxin from the spiderNephila clavata), were more potent antagonists of responses elicited bykainate (100 μM) or AMPA (30 μM). On the other hand, arylalkylamineswithin the two structural classes defined by Compound 1 and by Compound2 were found to inhibit preferentially responses evoked by NMDA (showingabout a 100-fold difference in potency). Thus, arylalkylamines such asCompound 1 and Compound 2 are potent and selective inhibitors of NMDAreceptor-mediated responses in cerebellar granule cells.

EXAMPLE 3 Patch Clamp Electrophysiology Studies

Patch clamp electrophysiological studies on isolated cortical orhippocampal neurons from adult rat brain have provided additionalinsight into the mechanism of action of Compound 1, Compound 2 andCompound 3. These studies revealed potent and selective inhibitoryeffects of arylalkylamines on responses mediated by NMDA receptors.Thus, compounds such as Compound 1 blocked responses to NMDA atnanomolar concentrations without affecting the responses to kainate.These results, which show selective inhibitory effects of thearylalkylamines in cortical and hippocampal neurons, indicate that thearylalkylamines target NMDA receptors in different regions within themammalian CNS. Moreover, it was found that the inhibitory effects ofthese compounds were use- and voltage-dependent. This strongly suggeststhat these compounds are blocking the open channel and, by this action,behave as noncompetitive NMDA receptor antagonists. Importantly,however, the arylalkylamines could be distinguished from both Mg²⁺ andMK-801, especially with respect to the voltage-dependence of their onsetof action and reversibility of effect.

EXAMPLE 4 Radioligand Binding Assays

Radioligand binding studies have demonstrated that arylalkylamine suchas Compound 1 and Compound 2 have a unique site of action. Although theyact like MK-801 in some respects (noncompetitive open-channel blockade,discussed above), they fail to displace [³H]MK-801 binding atconcentrations that completely block NMDA receptor-mediated responses.Assays such as these also demonstrate that the arylalkylamines do notbind with high affinity to the known MK-801, Mg²⁺ or polyamine bindingsites on the NMDA receptor-ionophore complex. Neither do thearylalkylamines bind directly to either the glutamate, glycine or esigmabinding sites at concentrations that block NMDA receptor-mediatedresponses. [³H] Compound 2 was synthesized as a radioligand for use inbinding studies to further explore the mechanism of action of Compound 2and particularly for use in a high-throughput screen to assess theactivity of other analogs and to detect new lead structures. A similarapproach was taken for [³H] COMPOUND 5. It is clear that compounds likeCompound 1 and Compound 2 target a site on the NMDA receptor-ionophorecomplex for which no other known compounds presently exist. The novelsite of action of the arylalkylamines at the molecular level translatesinto pronounced therapeutic advantages at the behavioral level. Asdescribed below, the arylalkylamines possess a quite differentbehavioral profile from other noncompetitive antagonists of the NMDAreceptor.

EXAMPLE 5 Synaptic Transmission Studies

The above findings demonstrate that certain arylalkylamines,specifically those related in structure to Compound 1 and Compound 2,act through a novel mechanism and site of action to potently andselectively inhibit NMDA receptor-mediated responses on neurons fromseveral different brain areas. To further assess the selectiveinhibitory actions of the arylalkylamines, their effects on synaptictransmission mediated by NMDA or AMPA receptors were assessed.

Glutamate-mediated transmission at synapses of Schaffer collateralfibers and CAl pyramidal cells was measured in slices of rat braincontaining the hippocampus. This assay measures electrophysiologicallythe postsynaptic depolarization caused by the presynaptic release ofglutamate, and can readily distinguish synaptic transmission mediated byNMDA or AMPA receptors. Arylalkylamines like Compound 1, Compound 2 andCompound 3 were again found to exert preferential inhibitory effects onNMDA receptor-mediated responses, and depressed responses mediated byAMPA receptors only at much higher concentrations. For example, Compound1 had an IC₅₀ for the NMDA receptor-mediated response of 20 μM, but anIC₅₀ for the AMPA receptor-mediated response of 647 μM. These resultsshow that arylalkylamines can selectively inhibit synaptic transmissionmediated by NMDA receptors other naturally occurring arylalkylaminespresent in the venom of Agelenopsis aperta likewise exert potent andselective inhibitory effects on NMDA receptor-mediated responses in therat hippocampus.

In the aggregate, then, the results of these various studies arecomplementary and together identify a structurally novel class ofcompounds with potent and selective inhibitory activity on NMDAreceptors in the mammalian CNS. Additionally, these compounds target aunique site on the NMDA receptor-ionophore complex. Compound 1, Compound2 and Compound 3 were selected for additional study in a variety of invitro and in vivo assays that model therapeutically important endpoints.

Neuroprotectant Activity

Desired properties of a neuroprotectant drug include the following. (1)The drug can be administered by oral or injectable routes (i.e., it isnot significantly broken down in the stomach, intestine or vascularsystem and thus reaches the tissues to be treated in a therapeuticallyeffective amount). Such drugs are easily tested in rodents to determinetheir bioavailability. (2) The drug exhibits neuroprotectant activity(i.e., efficacy) when given after an ischemic insult (stroke, asphyxia)or traumatic injury (head trauma, spinal cord injury). (3) The drug isdevoid of or has minimal side effects such as impairment of cognition,disruption of motor performance, sedation or hyperexcitability, neuronalvacuolization, cardiovascular activity, PCP-like abuse potential, orPCP-like psychotomimetic activity.

Although glutamate is the physiological synaptic transmitter, chronicexposure to glutamate leads to neuronal cell death. Much of theneurodegeneration caused by glutamate appears to be mediated by NMDAreceptors and results directly from chronically elevated levels ofcytosolic Ca²⁺. There is now extensive experimental support for the viewthat NMDA and AMPA receptors play a major role in mediating the neuronaldegeneration following a stroke and other ischemic/hypoxic events (Choi,Glutamate neurotoxicity and diseases of the nervous system. Neuron 1:623, 1988). Most of this evidence is based on the ability of competitiveor noncompetitive antagonists of the NMDA or AMPA receptor toeffectively block neuronal cell death in both in vitro and in vivomodels of stroke. Compound 1, Compound 2 and Compound 4 were thereforeexamined for neuroprotectant effects in standard assays designed todetect such activity.

EXAMPLE 6 Cortical Neuron Protection

To assess the in vitro neuroprotectant effect of arylalkylamines, mousecortical neurons grown in culture were exposed for 5 minutes to NMDA,and cell death after 24 hours was monitored by measuring the release oflactate dehydrogenase. (LDH), a cytoplasmic enzyme that is released fromdying cells (Choi et al., Glutamate neurotoxicity in cortical cellculture. J. Neurosci. 7: 357, 1987). Exposure to NMDA killed about 80%of the cortical neurons. Compound 1 or Compound 2, included along withNMDA, prevented cell death with IC₅₀ values of 70 μM and 30 μM,respectively. The effective concentrations of the arylalkylamines arehigher than those of other noncompetitive NMDA receptor antagonists, butsimilar to those of competitive antagonists. The effectiveconcentrations of NMDA receptor antagonists vary depending on theparticular experimental conditions and the type of cell studied‘cortical, hippocampal, striatal). This neuroprotectant effect likelyresults from the ability of these compounds to block the influx ofextracellular Ca²⁺ triggered by the NMDA receptor.

More rigorous testing to determine potential therapeutic efficacyinvolved in vivo stroke models. In these models, the blood supply istemporarily blocked by clamping the main arteries to the brain. Two invivo models of this sort were used to determine the ability of Compound1, Compound 2 and Compound 4 to prevent neuronal cell loss.

EXAMPLE 7 Bilateral Carotid Artery Occlusion

The first assay was the bilateral common carotid artery occlusion modelof forebrain ischemia performed in the gerbil (Karpiak et al., Animalmodels for the study of drugs in ischemic stroke. Ann. Rev. Pharimacol.Toxicol. 29: 403, 1989; Ginsberg and Busto, Rodent models of cerebralischemia. Stroke 20: 1627, 1989). Blood flow to the brain wasinterrupted for 7 minutes by clamping the carotid arteries. The testcompounds were administered as a single dose given intraperitoneally(i.p.) 30 minutes after reinstating blood flow. During the course ofthese experiments, the core body temperature of the animals wasmaintained at 37° C. to prevent any hypothermic reaction. It has beenshown that many NMDA receptor antagonists cause hypothermia and thiseffect can account for much of the protective effect of these compounds.The brains were examined for neuronal cell death 4 days later by silverstaining sections of the brain and quantifying death by morphometricanalysis. Compound 2 (20 mg/kg.) significantly (p<0.05) protectedagainst neuronal cell death in all areas of the brain examined (regionCAl of hippocampus, striatum and neocortex). Doses as low as 1 mg/kg.afforded complete (>98%) protection of the striatum. The degree ofprotection is comparable to that achieved with similar doses of thenoncompetitive NMDA antagonist, MK-801.

In subsequent experiments, Compound 1 (10 mg/kg) produced a 23%reduction in the amount of neuronal death in region CAl of the gerbilhippocampus measured at 7 days post-ischemia, while Compound 4 (10mg/kg) provided 90% protection.

EXAMPLE 8 Middle Cerebral Artery Occlusion

The middle cerebral artery model of stroke performed in the rat (Karpiaket al., Animal models for the study of drugs in ischemic stroke. Ann.Rev. Pharmacol. Toxicol. 29: 403, 1989; Ginsberg and Busto, Rodentmodels of cerebral ischemia. Stroke 20: 1627, 1989) is different,fromthe gerbil model because it results in a more restricted brain infarct,and thereby approximates a different kind of stroke—(focal thromboticstroke). In the first study using this stroke model, one cerebral arterywas permanently occluded by surgical ligation. The test compounds wereadministered 30 minutes after the occlusion by a single intraperitoneal(i.p.) injection. During the course of these experiments, the core bodytemperature of the animals was maintained at 37° C. to prevent anyhypothermic reaction. Brains were assessed histologically for neuronalcell loss 24 hours later. Infarct volumes were calculated using the areaof histological pallor from 10 slides and integrating the distancebetween each successive section. A single dose (30 mg/kg) of Compound 1was found to significantly (p<0.05) protect against neuronal cell lossequally as well as a maximally effective dose (10 mg/kg) of MK-801(approximately 15% protection). Preliminary studies with Compound 2 (20mg/kg) indicated a similar trend.

In the second study of focal cerebral ischemia in the rat, the middlecerebral artery was permanently occluded by passing a small piece ofsuture thread through the carotid artery to the region of the middlecerebral artery. Core body temperature was maintained at 37° C. Compound4, 10 mg/kg i.p. administered immediately after the onset of theischemic event, produced a statistically significant reduction in thevolume of the brain infarct (20%) recorded 24 hr later.

In a third model of focal cerebral ischemia in the rat, an ischemicinfarct was produced by a photothrombotic method using the dye RoseBengal. Compound 4, 10 mg/kg i.p. administered 30 min after the ischemicevent, produced a 20% reduction in the volume of the infarct, similar tothat seen with the noncompetitive NMDA receptor antagonist, MK-801.

In a fourth model of focal cerebral ischemia in the rat, the middlecerebral artery was temporarily occluded by passing a small piece ofsuture thread through the carotid artery to the region of the middlecerebral artery. The suture thread was withdrawn after an ischemicperiod of 2 hr. Core body temperature was maintained at 37° C. Compound4 administered at 10 mg/kg i.p. immediately after the onset of theischemic event, produced a statistically significant reduction in thevolume of the brain infarct (37%) recorded 72 hr later.

Several important features of the lead compounds emerge from these invivo results. First, and most importantly, Compound 1, Compound 2 andCompound 4 demonstrate neuroprotectant effects in several different invivo models of stroke. The gerbil assay is a model for transient globalcerebral ischemia and hypoxia such as cardiac arrest or perinatalhypoxia. The rat assays are models of permanent and temporary focalcerebral ischemia. The finding that Compound 1 and Compound 4 areneuroprotective in the permanent focal stroke models is surprisingbecause the accessibility of the drug to the site of infarction islimited to the penumbral region which generally is not large.Nonetheless, Compound 1 and Compound 4 significantly (p<0.05) limitedthe extent of damage. Second, the compounds are effective whenadministered after the ischemic event. This is important because thereis believed to be a “window of opportunity” following an infarct duringwhich drugs may effectively halt necrotic damage. How long this time isin humans has not been defined precisely, and will likely vary dependingupon the type of infarct. The essential observation, however, is thatthese compounds can prevent the spread of neuronal cell death once thedegenerative process has commenced. Finally, Compounds 1, 2, and 4 areeffective when administered parenterally, demonstrating that theypenetrate the blood-brain barrier.

Anticonvulsant Activity

Desired properties of an anticonvulsant drug include: the drug can beadministered by oral or injectable routes, the drug exhibits effectiveanticonvulsant activity against several seizure types, including, butnot limited to, simple partial seizures, complex partial seizures,status epilepticus, and trauma-induced seizures such as occur followinghead injury, including head surgery; and the drug is devoid of or hasminimal side effects such as impairment of cognition, disruption ofmotor performance, sedation or hyperexcitability, neuronalvacuolization, cardiovascular activity, PCP-like abuse potential, orPCP-like psychotomimetic activity.

Glutamate is the major excitatory transmitter in the brain, and thus mayplay a major role in seizure activity, and contribute to thepathogenesis of epilepsy. Much of the evidence favoring a major role forglutamate receptors in epilepsy derives from pharmacological studiesdemonstrating that glutamate receptor agonists elicit seizures, and thatNMDA and AMPA receptor antagonists are effective anticonvulsants whenadministered in vivo. There are numerous in vivo models involvingdifferent kinds of seizures and behavioral effects that are relevant forclinically distinct forms of epilepsy. It is thus prudent to test foreffects in several models, because it may be an oversimplification tosuppose that the same mechanism underlies all forms of seizure activity.

EXAMPLE 9 Convulsant Blocking Activity

In initial studies, the ability of arylalkylamines to block seizuresinduced by kainate, picrotoxin or bicuculline were examined. Each ofthese convulsants acts through a different mechanism and seizureselicited by kainate are qualitatively different from those elicited bypicrotoxin or bicuculline. In these experiments, a fraction ofAgelenopsis aperta venom containing several arylalkylamine toxins wasadministered intravenously (iv) 5 min before picrotoxin or bicuculline,and 5 min after kainate administration. The arylalkylamines diminishedthe seizures induced by all three of these agents. The effects ofpicrotoxin or bicuculline were so severe that all 19 control animalsdied within 25 minutes. In contrast, there were no deaths in the 9animals pretreated with the arylalkylamines. In fact, only about halfthe animals treated with the arylalkylamines showed any convulsions atall and those symptoms abated within an hour. These results demonstrateclear anticonvulsant effects of arylalkylamines and prompted furtherstudies using purified arylalkylamines and their analogs.

EXAMPLE 10 Seizure Stimuli

Three different seizure-inducing test paradigms were used initially inthis second group of studies and arylalkylamines such as Compound 1proved to be effective anticonvulsants in two such paradigms. The firsttwo models used DBA/2 mice which are prone to audiogenic seizures.Seizures were elicited by sound (bell tone at 109 dBs) or theintraperitoneal (ip) administration of NMDA (56 mg/kg). The testsubstances were administered 15-30 min before the convulsant stimulus.The number of clonic seizures was recorded for 1 min following theaudiogenic stimulus or for 15 min following the administration of NMDA.Compound 1, Compound 2, and several other arylalkylamines such asCompound 3 and Compound 4 depressed seizures evoked by either stimulus.For example, Compound 2 had an ED₅₀ of 0.13 mg/kg s.c. for audiogenicstimulus and 0.083 mg/kg s.c. for NMDA stimulus. Similarly, the EC₅₀ forCompound 4 in the audiogenic seizure model (0.08 mg/kg) approached thatfor MK-801 (0.02 mg/kg). In contrast, neither Compound 1 nor Compound 2was effective at doses up to 50 mg/kg s.c. in reducing seizures in CF-1mice elicited by i.p. NMDA.

In a second independent series of experiments, Compound 1 and Compound 4were found to prevent seizures induced by sound in another geneticallysusceptible mouse model of reflex epilepsy (Frings mice) followingintraperitoneal injection with IC₅₀ values of 14.3 mg/kg and −15 mg/kg,respectively. These compounds were considerably more potent againstaudiogenic seizures in Frings mice following intracerebro-ventricular(i.c.v.) injection, with IC₅₀ values of 0.63 μg (Compound 1) and 4.77 μg(Compound 4). Compound 1 was also found to be effective against seizureselicited by maximal electroshock in CF1 mice at a dose of 4 μg i.c.v.

In further studies using the genetically susceptible mouse model ofreflex epilepsy (Frings mice), Compound 9, Compound 12 and Compound 14,administered by i.c.v. injection, prevented sound-induced seizures withIC₅₀ values of 4.77 μg, 12.2 μg and 13.9 μg, respectively.

These collective findings demonstrate that arylalkylamines such asCompound 1, Compound 2 and Compound 4 are effective in preventingepileptic (audiogenic) and nonepileptic (chemoconvulsant) seizures. Thisgeneralized pattern of activity suggests that arylalkylamines areclinically useful in controlling seizure activity. In addition, thepotency of Compound 1, Compound 2 and especially Compound 4 in in vivomodels of seizure activity shows that these compounds can have thetherapeutically relevant effects when administrated parenterally in lowdoses, and are especially potent when administered directly into thecerebral ventricles.

Analgesic Activity

Desired properties of an analgesic drug include: the drug can beadministered by oral or injectable routes, the drug exhibits analgesicactivity, the drug is devoid of or has minimal side effects such asimpairment of cognition, disruption of motor performance, sedation orhyperexcitability, neuronal vacuolization, cardiovascular activity,PCP-like abuse potential, or PCP-like psychotomimetic activity.

Glutamate and NMDA receptor-mediated responses may play a role incertain kinds of pain perception (Dickenson, A cure for wind up: NMDAreceptor antagonists as potential analgesics. Trends Pharmacol. Sci. 11:302, 1990). The possible analgesic effects of Compound 1, Compound 2,Compound 3 and Compound 4 were therefore examined.

EXAMPLE 11 Writhing Response Test

In the first series of experiments, the animals were administered anunpleasant stimulus (2-phenyl-1,4-benzoquinone, PBQ) which elicits awrithing response (abdominal stretching). Typically, the number ofwrithes occurring in a 5 min observation period is recorded. Classicanalgesic drugs, such as morphine, are effective at decreasing thenumber of PBQ-elicited writhes (100% block of the writhing response at 4mg/kg i.p.). Nonsteroidal anti-inflammatory agents are likewiseeffective in this model. Compound 1 (2 mg/kg), Compound 2 (2 mg/kg) andCompound 3 (I mg/kg) depressed the writhing response by greater than 95%when administered s.c. or i.p. 30 minutes before PBQ. These resultsdemonstrate that Compound 1, Compound 2 and Compound 3 alleviatevisceral pain.

In a similar series of studies, Compound 1 and Compound 4 were found toinhibit acetic acid-induced writhing in mice following i.p. injectionwith IC₅₀ values of 10 mg/kg and 1 mg/kg, respectively.

EXAMPLE 12 Hot Plate Test

Compound 1 was tested for analgesic activity in an additional assay inthis model of analgesic activity, mice were administered test substancess.c. 30 min before being placed on a hot plate (50° C.). The time takento lick the feet or jump off the plate is an index of analgesicactivity, and effective analgesics increase the latency to licking orjumping morphine (5.6 mg/kg) increased the latency to jump by 765%.Compound 1 was likewise effective in this assay and, at doses of 4 and32 mg/kg, increased the latency to foot licking by 136% and the latencyto jumping by 360%, respectively.

It is noteworthy that the analgesic effects of Compound 1 in the hotplate assay were not accompanied by a decreased performance in theinverted grid assay (see below). This shows that the increase in thelatency to jump off the hot plate does not simply reflect impaired motorcapabilities. Together, these data suggest that Compound 1 possessessignificant analgesic activity.

In a later series of experiments, Compound 1 and Compound 4 weredemonstrated to possess significant analgesic activity in rats whenadministered by the intrathecal (i.th.) route. In these experiments, a52° C. hot plate was used as the nociceptive stimulus. Compound 1 (0.3-3nmol) and Compound 4 (0.3-3 nmol) produced dose- and time-dependentantinociceptive effects; these arylalkylamines were similar to morphine(0.3-3 nmol) in terms of potency and efficacy. The NMDA receptorantagonist, MK-801, on the other hand, was ineffective in this assay(3-30 nmol).

EXAMPLE 13 Tail Flick Test

In this standard assay, the thermal nociceptive stimulus was 52° C. warmwater with the latency to tail flick or withdrawal taken as theendpoint. Compound 1 (0.3-3 nmol) and Compound 4 (0.3-3 nmol) produced adose- and time-dependent analgesic effect following i.th.administration. These arylalkylamines were similar to morphine (0.3-3nmol) in terms of potency and efficacy. The NMDA receptor antagonist,MK-801, on the other hand, was ineffective in this assay (3-30 nmol).

EXAMPLE 14 Formalin Test

Male Sprague-Dawley rats were habituated to an observation chamber forat least 1 hr before receiving an injection of dilute formalin (5%) in avolume of 50 μl into the left rear paw. Behavioral responses weremonitored immediately after s.c. injection of formalin into the dorsalsurface of the paw by counting the number of flinches exhibited by theanimal. Behaviors were monitored for at least 50 min after formalininjection and were recorded as early phase responses (O-10 minpost-formalin) and late phase responses (20 50 min post-formalin).Compounds were injected intrathecally (i.th.) 10 min prior to formalin(pretreatment) or 10. min after formalin (post-treatment) in a volume of5 μl.

Intraplantal administration of formalin produced a typical biphasicresponse of flinching behavior, commonly described as the early and latephase responses. Intrathecal administration of Compound 1 (0.3-10 nmol)or Compound 4 (0.3-10 nmol) given as pretreatment to formalineffectively inhibited both early- and late-phase flinching behaviors.This effect of pretreatment with the arylalkylamines was, similar tothat seen with pretreatment with morphine (I-10 nmol) or MK-801 (1-30nmol).

Compound 1 (0.3-10 nmol i.th.) administered after the formalin producedsome inhibition of late-phase flinching, though significance wasachieved only at the 10 nmol dose. Compound 4 (0.3-10 nmol i.th.)administered after the formalin produced significant inhibition oflate-phase flinching, with significance achieved at the 3 and 10 nmoldoses. This analgesic profile of activity of the arylalkylamines issimilar to that seen with post-formalin administration of morphine (1-10nmol); post-formalin administration of MK-801 (1-30 nmol), however,failed to affect late-phase flinching.

Taken together, the results obtained with the hot plate, tail flick andformalin assays demonstrate that arylalkylamines such as Compound 1 andCompound 4 have significant analgesic activity in several rodent modelsof acute pain. The formalin assay additionally demonstrates thatarylalkylamines are effective in an animal model of chronic pain.Importantly, the arylalkylamines possess significant analgesic activitywhen administered after the formalin stimulus. This profile of activityclearly distinguishes the arylalkylamines from standard NMDA receptorantagonists such as MK-801.

Side Effects of Arylalkylamines

Given the important role NMDA receptors play in diverse brain functions,it is not surprising to find that antagonists of this receptor aretypically associated with certain unwelcome side effects. In fact, it isthis property that provides the major obstacle to developing therapiesthat target NMDA receptors. The principal side effects,which-characterize both competitive and noncompetitive antagonists, area PCP-like psychotomimetic activity, impairment of motor performance,sedation or hyperexcitability, impairment of cognitive abilities,neuronal vacuolization, or cardiovascular effects (Willetts et al., Thebehavioral pharmacology of NMDA receptor antagonists. Trends Pharmacol.Sci. 11: 423, 1990; Olney et al., Pathological changes induced incerebrocortical neurons by phencyclidine and related drugs. Science 244:1360, 1989). The psychotomimetic effect associated with inhibition ofNMDA receptor-mediated responses is epitomized in the response tophencyclidine (PCP) or “angel dust” which acts at the MK-801 bindingsite. Impairment of cognitive ability is associated with the importantrole that NMDA receptors normally play in learning and memory.

Relatively less is known concerning the side effect profile of AMPAreceptor antagonists. However, it is becoming clear that such compoundsalso elicit motor impairment, ataxia and profound sedation.

The activity of arylalkylamines was examined in animal models that indexmotor impairment, sedation and psychotomimetic activity as well as bothin vitro and in vivo models of learning and memory.

(a) PCP-Like Psychotomimetic Activity

In rodents, both competitive and noncompetitive antagonists of the NMDAreceptor produce a PCP-like stereotypic behavior characterized byhyperactivity, head-weaving, and ataxia (Willetts et al., The behavioralpharmacology of NMDA receptor. antagonists. Trends Pharmacol. Sci. 11:423, 1990; Snell and Johnson, In: Excitatory Amino Acids in Health andDisease, John Wiley & Sons, p. 261, 1988). We investigated whether thearylalkylamines would elicit such behaviors. In addition, weinvestigated whether the arylalkylamines would substitute for PCP inrats trained to discriminate PCP from-saline (Willetts et al., Thebehavioral pharmacology of NMDA receptor antagonists. Trends Pharmacol.Sci. 11: 423, 1990), and whether the arylalkylamines would elicit aPCP-like neuronal vacuolization (Olney et al., Pathological changesinduced in cerebrocortical neurons by phencyclidine and related drugs.Science 244: 1360, 1989).

EXAMPLE 15 Locomotor Activity

The first assay simply monitors locomotor activity during the first hourfollowing peripheral (s.c. or i.p.) administration of test substance.Mice received a dose of Compound 1 15 min before being placed intoactivity chambers. Activity was quantified by counting the number ofbreaks in a phototube grid in a 60 min period. In this assay, MK-801(0.25 mg/kg p.o.) causes a 2- to 3-fold increase in locomotor activity.However, Compound 1, even when tested at 32 mg/kg s.c., did not elicithyperactivity and, in fact, tended to depress it. This result, using apurified arylalkylamine in mice, complements earlier results obtained-inrats where the entire arylalkylamine-containing fraction fromAgelenopsis aperta, when injected intravenously, did not elicit aPCP-like behavioral syndrome but seemed to produce a mild sedativeeffect.

EXAMPLE 16 Motor Impairment

In the first assay for generalized motor impairment, Compound I wasexamined-in the inverted grid assay. In this assay, animals are placedon a wire-holed grid suspended from a rotating metal bar which can beinverted. The animals are then scored for their ability to climb to thetop or hang on to the grid. Animals with severe motor impairment falloff the grid. This assay provides an index of “behavioral disruption”that may result from ataxia, loss of the righting reflex, sedation, ormuscle relaxation. In these tests, Compound 1, administered at 32mg/kg.s.c., did not lessen the ability of DBA/2 mice to right themselveswhen the grid was inverted (p>0.05). Compound 2 was likewise withouteffect (p>0.05) on motor performance in DBA/2 mice when administered ata dose of 20 mg/kg s.c. These doses are considerably higher than thoserequired to prevent sound-induced seizures in DBA/2 mice (see Example 10above).

The second assay of acute motor impairment was the rotorod assay. Inthis assay, Frings and CF1 mice were injected with test compound andplaced on a knurled rod which rotated at a speed of 6 rpm. The abilityof the mice to maintain equilibrium for long periods of time wasdetermined; those mice that were unable to maintain equilibrium on therotorod for 1 min in each of 3 trials were considered impaired. Compound1 produced acute motor impairment in Frings mice with a TD, (that dosewhich produced motor toxicity in 50% of the test animals) of 16.8 mg/kgi.p. This dose is similar to that which prevents sound-induced seizuresin Frings mice (see Example 10 above). There is a much clearerseparation between effective and toxic doses of Compound 1 in Fringsmice, however, when the Compound is administered i.c.v. In this case, noapparent motor toxicity was evident until the dose of Compound 1exceeded 1.56 μg i.c.v. (>2 times the ED₅₀ of 0.63 μg). Finally, motorimpairment in @—F1 mice was noted with Compound 1 following i.c.v.administration of 4 μg.

Compound 4, Compound 9, Compound 12 and Compound 14 were administered toFrings mice by i.c.v. injection, and acute motor impairment wasmeasured. The TD₅₀ values for Compounds 4, 9, 12 and 14 were 8-16 μg,14.8 μg, 30.2 μg and 30.8 μg, respectively. These TD₅₀ values were 2-3times higher than the effective IC₅₀ values for anticonvulsant potency(see Example 10 above); a clear separation between effective and toxicdoses was noted.

EXAMPLE 17 PCP Discrimination

In this assay, rats who have been trained to lever press for foodreinforcement must select which of two levers in their cages is correct.The only stimulus they have for selecting the correct lever is theirability to detect whether @hey received a PCP or vehicle injection.After about two months of training, rats become very good atdiscriminating PCP from vehicle injections and can then be tested withother drugs to determine if they are discriminated as PCP. When testedin this procedure, other drugs which are known to produce a PCP-likeintoxication substitute for PCP. These drugs include various PCP analogssuch as ketamine and the noncompetitive NMDA receptor antagonist,MK-801.

Compound 1 (1-30 mg/kg i.p.) did not substitute for PCP, and thus wascompletely devoid of PCP-like discriminative stimulus effects. At 30mg/kg i.p., only 1 of the 7 animals tested responded at all on eitherlevel. It is thus clear that a behaviorally effective dosage range ofCompound 1 was evaluated. As the ability of test compounds to producePCP-like effects in rats is believed to be predictive of their abilityto produce PCP-like psychotomimetic activity and abuse liability inhumans, these results strongly suggest that the arylalkylamines such asCompound 1 will lack such deleterious side effects in man.

EXAMPLE 18

The administration of compounds such as PCP and MK-801 to rats producesa neurotoxic effect termed neuronal vacuolization. Following a singledose of such compounds, vacuoles are found in particular centralneurons, especially those in the cingulate cortex and retrosplenialcortex. No such vacuolization was present in-rats treated with CompoundI at the single high dose of 100 mg/kg i.p.

Taken together, the results on locomotor activity, motor impairment, PCPdiscrimination and neuronal vacuolization strongly suggest thatarylalkylamines will be devoid of PCP-like side effects in man.

(b) Cognitive Impairment

One of the major reasons for postulating a role of NMDA receptors inmemory and learning derives from cellular studies on long-termpotentiation (LTP) in the rat hippocampus. LTP is a long-lastingincrease in the magnitude of synaptic responses produced by brief yetintense synaptic stimulation. Since the discovery of this phenomenon, ithas become the preeminent cellular model of learning in the vertebratebrain (Teyler and Discenna, Long-term potentiation. Annu. Rev. Neurosci.10: 131, 1987). Transmission at synapses formed by Schaffer collateralsonto CAl pyramidal cells is mediated by NMDA and AMPA receptors.Following a brief tetanizing stimulus, the magnitude of the populationspike (a measure of synaptic transmission) is greatly increased andremains so for hours. It has been shown that all known competitive andnoncompetitive antagonists of NMDA receptors block LTP in the rathippocampus, whereas antagonists of non-NMDA receptors are withouteffect (Collingridge and Davis, In: The NMDA Receptor, IRL Press. p.123, 1989). This supports a role of NMDA receptors in memory andlearning.

EXAMPLE 19 LTP Assay

The effects of selected arylalkylamines and literature standards wereexamined for effects on LTP in slices of rat hippocampus. Asanticipated, all the conventional competitive (AP5 and AP7) andnoncompetitive (MK-801 and ifenprodil) NMDA receptor antagonistsinhibited the induction of LTP in the hippocampus. Slices of rathippocampus were superfused for 30-60 min with a test compound beforedelivering a tetanizing stimulus consisting of 3 trains, separated by500 msec, of 100 Hz for 1 sec each. The response amplitude was monitoredfor an additional 15 minutes post-tetanus. The tetanizing stimuluscaused a mean 95% increase in the amplitude of the synaptic response.The induction of LTP was significantly blocked (p<0.05) by competitive(AP5, AP7) or noncompetitive (MK-801, ifenprodil) NMDA receptorantagonists. Quite surprisingly, none of the arylalkylamines tested(Compound 1, Compound 2, Compound 3 and others) blocked the induction ofLTP (p>0.05), even when used at high concentrations (100-300 μM) thatcaused some inhibition of the control response.

These results highlight yet another unique and important feature ofarylalkylamines. Arylalkylamines are the first, and at present the only,class of compounds shown to be selective and potent antagonists of theNMDA receptor that do not block the induction of LTP. This likelyreflects the novel mechanism and site of action of arylalkylamines andsuggests that drugs which target the novel site on the NMDA receptorwill similarly lack effects on LTP As LTP is the primary cellular modelfor learning and memory in the mammalian CNS, it additionally suggeststhat such drugs will lack deleterious effects on cognitive performance.

EXAMPLE 20 Learning Tests

Preliminary experiments using one of the more potent syntheticarylalkylamine analogs, Compound 3, in an in vivo learning paradigmdemonstrate that these drugs lack effects on cognitive performance. Inthis test, rats were trained to alternate turning in a T maze for a foodreward. MK-801 was included for comparison. Test compounds wereadministered i.p. 15 min before testing. Control animals made thecorrect choice about 80% of the time. Increasing doses of MK-801progressively decreased the number of correct choices and this decrementin behavior was accompanied by hyperactivity. In contrast, Compound 3did not impair the ability of the animals to make the correct choices(p>0.05). At the highest doses tested, Compound 3 caused some decreasein locomotor activity, exactly the opposite effect observed with MK-801.

Although MK-801 decreased learning performance in parallel withincreases in locomotor activity, other studies using different paradigmsin rodents and primates have shown a clear dissociation between theeffects on learning and locomotion. Thus, both competitive andnoncompetitive NMDA receptor antagonists impair learning at doses whichdo not cause any overt change in motor behavior. This demonstrates thatconventional NMDA receptor antagonists impair learning independently ofother side effects. The results of the T-maze assay demonstrate thatCompound 3, and other arylalkylamines, do not impair learning even atdoses that cause some decrease in locomotor activity.

One additional observation emerged from these learning tests. Theanimals, first response on the second day of testing was random and wastherefore not dependent on the last response of the previous day'stesting. Control animals thus correctly made the first choice about 50%of the time. MK-801 has no effect on this first choice. However, animalsadministered Compound 3 on the previous day made the first choicecorrectly considerably more often. Unlike control animals then, theanimals treated with Compound 3 behaved as if they remembered the lastchoice of the previous day.

In a second series of experiments, the effect of Compound 4 on learningin the Morris water maze task was evaluated. In this test, a hiddenplatform was placed in a fixed location in a circular steel tank, andsubmerged 2 cm below the surface of the water. Each rat was given 3trials per day with a 10 min intertrial interval for 5 days. A trial wasinitiated by placing the rat in the water, nose facing the wall of thetank, at one of three predetermined starting locations. The order of thestart location was varied daily. Learning was measured as a decrease intime required to swim to the platform. If an animal failed to locate theplatform within 60 sec after the start of the trial, the rat washand-guided to it. The animals remained on the platform for 10 secbefore being removed from the tank. Ten min after the last trainingtrial on day 5, the animals received a probe test. The platform wasremoved for this 1 trial task and the animals were allowed to swim for60 sec to assess the spatial bias for the platform location. Twomeasures were recorded from this task: latency to first crossing thearea where the platform had been, and total number of crossings. A totalof 5 injections of Compound 4 were given to each rat. In the firstseries of experiments, Compound 4 was administered at 10 mg/kg i.p.daily for 5 days. This treatment regimen impaired learning; however,these animals experienced significant weight loss and unusual behavioralsigns (“shivering,” motor impairment, difficulty in swimming) withrepeated dosing of Compound 4. In a subsequent study, six animalsreceived 1 mg/kg i.p. for the first 4 days of training, while twoanimals received 5 mg/kg i.p. during this period on the last day oftraining, both groups received 10 mg/kg. Neither the 1 mg/kg nor the 5mg/kg animals showed any impairment in learning the location of thehidden platform, nor did the final 10 mg/kg dose produce any impairmentin the ability of the animal to perform the already learned task.

The results of these learning tasks are encouraging. They suggest thatarylalkylamines lack the learning and memory deficits that typify otherNMDA receptor antagonists. In fact, there is a suggestion that thearylalkylamines may even be nootropic (memory enhancers).

(c) Cardiovascular Effects

In vivo studies with certain arylalkylamines revealed a hypotensiveeffect of these compounds, especially at high doses. On the basis ofthese results, a systematic study of the effects of arylalkylamines oncardiovascular function was performed.

EXAMPLE 21 Ca²⁺ Channel Inhibition

We have discovered that some of the arylalkylamines are quite potentinhibitors of voltage-sensitive Ca²⁺, channels, specifically thosesensitive to inhibition by dihydropyridines (L-type channels). Sucheffects on vascular smooth muscle would be expected to dilate bloodvessels and cause a drop in blood pressure, thus producing hypotension.

The ability of arylalkylamines to inhibit dihydropyridine-sensitive Ca²⁺channels was examined in cerebellar granule cells and a rat aorticsmooth muscle cell line, A₇r5 cells. In cerebellar granule cells,Compound 2 inhibited depolarization-induced increases in [Ca²⁺]_(I) atconcentrations 100-fold higher than those required to block responses toNMDA (IC50 values of 24 μM and 161 nM, respectively). Overall, we haveobserved a wide range of potencies against voltage-sensitive Ca²⁺channels that does not correlate with potency against NMDA receptors.This strongly suggests that further structure-activity work based onchemical modification of the arylalkylamine molecule will lead to thedevelopment of compounds that are very potent NMDA antagonists with lowpotency against voltage-sensitive Ca²⁺ channels. Indeed, Compound 1(with an IC₅₀ of 102 nM against NMDA receptor-mediated responses incerebellar granule cells) is a relatively poor inhibitor ofvoltage-sensitive Ca²⁺ channels in cerebellar granule cells (IC₅₀=257μM) and is virtually without effect on voltage-sensitive Ca²⁺ influx inA₇r5 cells (ICs₅₀=808 μM).

Arylalkylamines are not, however, indiscriminate blockers ofvoltage-sensitive Ca²⁺ channels. They do not, for example, affectvoltage-sensitive Ca²⁺ channels in cerebellar Purkinje cells (P-typechannels) or those channels thought to be involved in neurotransmitterrelease (N-channels). The arylalkylamines that do blockvoltage-sensitive Ca²⁺ channels appear to target specifically L-typeCa²⁺ channels. Moreover, as mentioned above, there is a high degree ofstructural specificity in this effect. For example, one arylalkylamineis 57 times more potent than another arylalkylamine in blocking Ca²⁺influx through L-type channels, where the only structural differencebetween the compounds is the presence or absence of a hydroxyl group.

EXAMPLE 22 In Vivo Cardiovascular Studies

The arylalkylamines Compound 1 and Compound 2 produce moderate drops(20-40 mm Hg) in mean arterial blood pressure (MABP) in anesthetizedrats at doses which are effective in the in vivo stroke models (10-30mg/kg s.c.). The hypotensive effect of Compound 4 has been evaluated ingreater detail. Compound 4 elicited a marked drop (40 mm Hg) in meanarterial pressure which persisted for approximately 90-120 min whenadministered at the dose of 10 mg/kg i.p.; it was in this same group ofrats that Compound 4 afforded significant neuroprotection in the suturemodel of middle cerebral artery occlusion (see Example 8 above). Similarresults were obtained in the rat study in which Compound 4 demonstratedneuroprotectant activity in the Rose Bengal photothrombotic model offocal ischemic stroke (see Example 8 above). Further studies using thepithed rat preparation strongly suggest that the hypotensive activity ofCompound 4 is a peripherally mediated effect. The hypotension andbradycardia produced by Compound 4 was maintained in rats pretreatedwith atropine, suggesting that these effects are not mediated by acholinergic mechanism. Similarly, Compound 4 elicited hypotension andbradycardia in chemically sympathectomized rats (pretreated with aganglionic blocker), suggesting that these effects are not mediated viathe sympathetic nervous system.

On the basis of these findings, it is anticipated that chemical effortswill minimize the cardiovascular side effects by (1) enhancing theuptake of arylalkylamine into the brain such that lower doses arerequired for neuroprotection, and (2) increasing the selectivity(potency ratio) of arylalkylamines for receptor-operated Ca²⁺ channelsover voltage-sensitive Ca²⁺ channels.

EXAMPLE 23 Biological Activity of Compound 19 and Analogs

Compounds 19-215 had high potencies against NMDA-induced increases in[Ca²⁺]_(I) rat cerebellar granule cells grown in culture (Table 1). Theinhibitory effect of Compound 19 on responses to NMDA wasnoncompetitive. Compounds 19-215. inhibited [³H]MK-801 binding inmembranes prepared from rat hippocampal and cortical tissue (Table 1).

Compound 19 possessed the following additional biological activities:significant (p<0.05 compared to control) anticonvulsant activity againstmaximal electroshock-induced seizures in mice following i.p.administration (ED₅₀ =26.4 mg/kg and TD₅₀ (rotorod)=43.8 mg/kg);significant anticonvulsant activity against maximal electroshock-inducedseizures in mice following oral (p.o.) administration (ED₅₀=35 mg/kg),but with motor impairment at 30 mg/kg; significant analgesic activity inthe hot-plate and PBQ-induced writhing assays at 16 mg/kg i.p.; noPCP-like stereotypic behavior (hyperexcitability and head weaving) at 30mg/kg i.p. in rats; no generalization to PCP in the PCP discriminationassay in rats at doses up to the behaviorally active dose of 30 mg/kgi.p. Compound 19 was significantly less potent in antagonizing increasesin [Ca²⁺]_(I) elicited by depolarizing concentrations of KCl in ratcerebellar granule cells (IC50=10.2 μM), and was without effect on bloodpressure when administered s.c. in rats at doses up to 100 mg/kg.Compound 19, however, did block the induction of LTP in rat hippocampalslices when tested at 100 μM.

Compound 20 possessed the following additional biological activities:significant anticonvulsant activity against maximal electroshock-inducedseizures in mice following i.p. administration (ED50=20.1 mg/kg and TD₅₀(rotorod)=20.6 mg/kg); no significant anticonvulsant activity againstmaximal electroshock-induced seizures in mice following oral (p.o.)administration at doses up to 30 mg/kg, but with motor impairment at 30mg/kg; significant anticonvulsant activity against sound-inducedseizures in a genetically susceptible mouse model of reflex epilepsy(Frings mice) following i.p. (ED₅₀=2.1 mg/kg and TD₅₀=19.9 mg/kg) andoral (ED₅₀=9.7 mg/kg and TD₅₀=21.8 mg/kg) administration; significantanticonvulsant activity against maximal electroshock-induced seizures inrats following oral administration with an ED₅₀ value of 33.64 mg/kg andan TD₅₀ value of 55.87 mg/kg; an increase in seizure threshold asindexed by the i.v. Metrazol test in mice at the dose of 10 mg/kg i.p.;significant neuroprotectant activity in a rat model of temporary focalischemia (a 51% reduction in the infarct volume following theadministration of two doses of 1 mg/kg i.p., the first given immediatelyafter middle cerebral artery occlusion and the second given 6 hr later;a 43% reduction in the infarct volume following the administration oftwo doses of 1 mg/kg i.p., the first given 2 hr after middle cerebralartery occlusion (i.e., at the time of reperfusion) and the second given6 hr later); significant neuroprotectant activity (a 24% reduction inthe infarct volume) in a rat model of permanent focal ischemia followingthe administration of 1 mg/kg i.p. at 30 min and again 4 hrpost-occlusion; significant neuroprotectant activity (a 50% reduction inthe infarct volume) in a rat photothrombotic model of focal ischemiafollowing tie administration of 10 mg/kg i.p. at 15 min, 3 hr, and again6 hr post-occlusion; no significant analgesic activity at the dose of 25mg/kg i.p. in the rat 52° C. hot plate test or the rat 48° C. tail flicktest; significant analgesic activity, not blocked by the opiate receptorantagonist naloxone, in the rat formalin test at the dose of 10 mg/kgi.p.; significant analgesic activity, not blocked by naloxone, againstacetic acid-induced abdominal writhing in mice at the dose of 10 mg/kgi.p.; no generalization to PCP in the PCP discrimination assay in ratsat doses up to the behaviorally active dose of 10 mg/kg i.p.; noneuronal vacuolization in rats when administered at doses of 10 and 30mg/kg i.p.; no significant cardiovascular activity in anesthetized ratsat doses up to 15 μmoles/kg i.v. or 10 mg/kg i.p.; no significantcardiovascular activity in conscious beagle dogs at doses of 0.3 or 1mg/kg i.v. (60 sec bolus injection); transient increases in meanarterial pressure and heart rate in conscious beagle dogs at the dose of3 mg/kg i.v., with larger magnitude and longer duration effects seen atthe dose of 10 mg/kg i.v. (60 sec bolus injection); increased motoractivity, agitation and anxiousness, slight tremors, licking of themouth, whining,and urination in conscious beagle dogs at the dose of 3mg/kg i.v. (60 sec bolus injection); dilated pupils, whole body tremors,incoordination, licking of the mouth, salivation, panting, rapidblinking of the eyes, whining, anxiousness, seizures, and death inconscious beagle dogs at the dose of 10 mg/kg i.v. sec bolus injection);no behavioral effects in conscious male NMRI mice at the doses of 2 and4 mg/kg i.p.; excitation and increased reactivity to touch in consciousmale NMRI mice at the dose of 8 mg/kg i.p.; excitation, Straub tail,tremor, stereotypies, hypothermia, and mydriasis in conscious male NMRImice at the doses of 16 and 32 mg/kg i.p.; convulsions and death inconscious male NMRI mice at the dose of 64 mg/kg i.p.; convulsion's anddeath in conscious male NMRI mice at the doses of 128 and 256 mg/kgi.p.; no behavioral effects in conscious male Wistar rats at the dose of2 mg/kg i.v.; excitation, stereotypies, increased reactivity to touch,increased muscle tone, and tremor in conscious male wistar rats at dosesranging from 4 to 16 mg/kg i.v.; Straub tail, convulsions, and death inconscious male wistar rats at the dose of 32 mg/kg i.v.

Compound 21 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀ =3.41 mg/kg and TD₅₀ (motorimpairment)=15.3 mg/kg).

Compound 33 (an enantiomer of Compound 21)possessed the followingadditional biological activities:significant anticonvulsant activityagainst sound-induced seizures in a genetically susceptible mouse modelof reflex epilepsy (Frings mice) following i.p. administration (ED₅₀=4.6mg/kg and TD₅₀ (motor impairment)=27.8 mg/kg); significantanticonvulsant activity against maximal electroshock-induced seizures inrats following oral administration at the dose of 25 mg/kg, with nomotor toxicity apparent at this dose; significant neuroprotectantactivity in a rat model of focal ischemic stroke following i.p.administration of 2 mg/kg 30 min prior to vessel occlusion and 2 mg/kg 3hr post-occlusion; no significant analgesic activity at the dose of 25mg/kg i.p. in the rat 52° C. hot plate test or the rat 48° C. tail flicktest; significant analgesic activity in a rat model of chronicneuropathic pain following i.th. administration of doses ranging from 15to 80 μg; significant analgesic activity in a rat model of chronicneuropathic pain following i.p. administration of doses of 3-10 mg/kg;no neuronal vacuolization when administered to rats at the dose of 30mg/kg i.p.; no significant cardiovascular activity in anesthetized ratsat doses up to 3 mg/kg i.v.; no significant cardiovascular activity inconscious beagle dogs at the dose of 0.3 mg/kg i.v. (60 sec bolusinjection); transient increases in mean arterial pressure in consciousbeagle dogs at the dose of 1 mg/kg i.v., with larger magnitude andlonger duration effects seen at the doses of 3 and 10 mg/kg i.v. (60 secbolus injection); a transient increase in heart rate in conscious beagledogs at the dose of 10 mg/kg.i.v. (60 sec bolus injection); licking ofthe mouth in conscious beagle dogs at the dose of 3 mg/kg i.v. (60 secbolus injection); dilated pupils, whole body tremors, incoordination,licking of the mouth, salivation, and panting in conscious beagle dogsat the dose of 10 mg/kg i.v. (60 sec bolus injection); no significantdrug-induced changes in the ECG in conscious beagle dogs at doses up to10 mg/kg i.v. (60 sec bolus injection); no behavioral effects inconscious male NMRI mice at the doses of 2 and 4 mg/kg i.p.; excitation,increased reactivity to touch, and hypothermia in conscious male NMRImice at the dose of 8 mg/kg i.p.; excitation, Straub tail, tremor,jumping, stereotypies, hypothermia, and mydriasis in conscious male NMRImice at the doses of 16 and 32 mg/kg i.p.; convulsions in conscious maleNMRI mice at the dose of 64 mg/kg i.p.; convulsions and death inconscious male NMRI mice at the doses of 128 and 256 mg/kg i.p.

Compound 34 (an enantiomer of Compound 21) possessed the followingadditional biological activities: significant anticonvulsant activityagainst sound-induced seizures in a genetically susceptible mouse modelof reflex epilepsy (Frings mice) following i.p. administration (ED₅₀=22mg/kg and TD₅₀ (motor impairment) between 10 and 15 mg/kg); hyperthermiain conscious male NMRI mice at the dose of 2 mg/kg i.p.; no behavioraleffects in conscious male NMRI mice at the dose of 4 mg/kg i.p.;excitation, increased reactivity to touch, and hypothermia in consciousmale NMRI mice at the dose of 8 mg/kg i.p.; excitation, Straub tail,tremor, jumping, stereotypies, hypothermia, and mydriasis in consciousmale NMRI mice at the doses of 16 and 32 mg/kg i.p.; convulsions inconscious male NMRI mice at the dose of 64 mg/kg i.p.; convulsions anddeath in conscious male NMRI mice at the doses of 128 and 256 mg/kg i.p.

Compound 22 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. (ED₅₀=4.9 mg/kg and TD₅₀, (inverted grid)=26.8 mg/kg) andoral (ED₅₀=5.1 mg/kg and LD₅₀-18.3 mg/kg) administration; and nosignificant cardiovascular activity in anesthetized rats at doses up to15 μmoles/kg (4.47 mg/kg) i.v.

Compound 50 (an enantiomer of Compound 22) possessed the followingadditional biological activities: significant anticonvulsant activityagainst sound-induced seizures in a genetically susceptible mouse modelof reflex epilepsy (Frings mice) following i.p. administration (ED₅₀=2.7mg/kg and TD₅₀ (motor impairment)=17.4 mg/kg); significantanticonvulsant activity against sound-induced seizures in a geneticallysusceptible mouse model of reflex epilepsy (Frings mice) following p.o.administration (ED₅₀=9.0 mg/kg and TD₅₀ (motor impairment)=18.9 mg/kg);significant anticonvulsant activity against maximal electroshock-inducedseizures in rats following oral administration with ED₅₀=28 mg/kg andTD₅₀=20 mg/kg; significant neuroprotectant activity in a rat model offocal ischemic stroke following i.p. administration of 2 mg/kg 30 minprior to vessel occlusion and 2 mg/kg 3 hr post-occlusion; nosignificant analgesic activity at the dose of 25 mg/kg i.p. in the rat52° C. hot plate test or the rat 48° C. tail flick test; and nosignificant cardiovascular activity in anesthetized rats at doses up to5 mg/kg i.v.

Compound 51 (an enantiomer of Compound 22) possessed the followingadditional biological activities: significant anticonvulsant activityagainst sound-induced seizures in a genetically susceptible mouse modelof reflex epilepsy (Frings mice) i.p. administration (ED₅₀=9.1 mg/kg andTD₅₀, (motor impairment)=13.6 mg/kg).

Compound 24 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=5 mg/kg and TD₅₀ (motorimpairment)=16 mg/kg); significant anticonvulsant activity againstmaximal electroshock-induced seizures in rats following oraladministration with ED₅₀=46 mg/kg and TD₅₀=51 mg/kg; no significantneuroprotectant activity in a rat model of focal ischemic strokefollowing i.p. administration of 2 mg/kg 30 min prior to vesselocclusion and 2 mg/kg 3 hr post occlusion; and no significantcardiovascular activity in anesthetized rats at doses up to 10 mg/kgi.v.

Compound 25 possessed the following additional biological activities:significant anticonvulsant activity against maximal electroshock-inducedseizures in mice following i.p. administration with an ED₅₀=mg/kg and aTD₅₀=32.18 mg/kg; significant anticonvulsant activity against maximalelectroshock-induced seizures in rats following oral administration withan ED50=46.43 mg/kg and a TD., between 163 and 326 mg/kg.

Compound 31 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=6 mg/kg and TD₅₀ (motor impairment)between 10 and 20 mg/kg).

Compound 46 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=25 mg/kg and TD₅₀ (motor impairment)between 18 and 21 mg/kg); and no significant neuroprotectant activity ina rat model of focal ischemic stroke following i.p. administration of 2mg/kg 30 min prior to vessel occlusion and 2 mg/kg 3 hr post-occlusion.

Compound 57 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=1 mg/kg and TD₅₀ (motor impairment)between 6 and 8 mg/kg).

Compound 58 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=0.9 mg/kg and TD₅₀ (motorimpairment)=14.5 mg/kg); no significant neuroprotectant activity in arat model of focal ischemic stroke following i.p. administration of 2mg/kg 30 min prior to vessel occlusion and 2 mg/kg 3 hr post-occlusion;and no significant cardiovascular activity in anesthetized rats at dosesup to 2 mg/kg i.v.

Compound 59 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=2.7 mg/kg and TD₅₀ (motorimpairment)=7.8 mg/kg); a reduction in seizure threshold as indexed bythe i.v. Metrazol test in mice at the dose of 11.7 mg/kg i.p.; nosignificant neuroprotectant activity in a rat model of focal ischemicstroke following i.p. administration of 2 mg/kg 30 min prior to vesselocclusion and 2 mg/kg 3 hr post-occlusion; and no significantcardiovascular activity in anesthetized rats at doses up to 10 mg/kgi.v.

Compound 60 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration (ED₅₀=4.4 mg/kg and TD₅₀ (motorimpairment)=9.2 mg/kg); significant anticonvulsant activity againstsound-induced seizures in a genetically susceptible mouse model ofreflex epilepsy (Frings mice) following oral administration (ED₅₀=10mg/kg and TD₅₀ (motor impairment)=25.6 mg/kg); significantanticonvulsant activity against maximal electroshock-induced seizures inmice following i.p. administration (ED₅₀=8.17 mg/kg and TD₅₀(rotorod)=17.30 mg/kg); no effect on seizure threshold as indexed by thei.v. Metrazol test in mice at the doses of 1 and 4 mg/kg i.p.; areduction in seizure threshold as indexed by the i.v. Metrazol test inmice at the doses of 8 and 17 mg/kg i.p.; significant neuroprotectantactivity in a rat model of temporary focal ischemic stroke followingi.p. administration of 2 mg/kg 30 min prior to vessel occlusion and 2mg/kg 3 hr post-occlusion; significant neuroprotectant activity in a ratmodel of temporary focal ischemic stroke following i.p. or i.v.administration of 1 mg/kg 2 hr and again 8 hr post-occlusion;significant neuroprotectant activity in a rat model of temporary focalischemic stroke following i.v. administration of 1 mg/kg 2 hrpost-occlusion; no significant neuroprotectant activity in a ratphotothrombotic model of focal ischemia following the administration of10 mg/kg i.p. at 15 min, 3 hr, and again 6 hr post-occlusion; noneuronal vacuolization when administered at, doses of 20 mg/kg i.p. or10 mg/kg i.v.; no significant cardiovascular activity in consciousbeagle dogs at the dose of 0.3 mg/kg i.v. (60 sec bolus injection);transient increases in mean arterial pressure in conscious beagle dogsat the doses of 1 and 3 mg/kg i.v., with larger magnitude and longerduration effects seen at the dose of 10 mg/kg i.v. (60 sec bolusinjection); transient increases in heart rate in conscious beagle dogsat the doses of 3 and 10 mg/kg i.v. (60 sec bolus injection), nosignificant changes in the ECG in conscious beagle dogs at doses rangingfrom 0.3 to 10 mg/kg i.v. (60 sec bolus injection); no significantbehavioral effects in conscious beagle dogs at the doses of 0.3 and 1mg/kg i.v. (60 sec bolus injection); a slight increase in respiratoryrate in conscious beagle dogs at the dose of 0.3 mg/kg i.v. (60 secbolus injection); dilated pupils, whole body tremors, salivation, andurination in conscious beagle dogs at the dose of 10 mg/kg i.v. (60 secbolus injection); no significant behavioral effects in conscious maleWistar rats at doses up to 4 mg/kg i.v.; excitation, stereotypies,increased reactivity to touch, increased muscle tone, and tremor inconscious male Wistar rats at the dose of 8 mg/kg i.v.; Straub tail,convulsions, and death in Unconscious male Wistar rats at the dose of 16mg/kg i.v.

Compound 119 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration with an ED₁₀=7.0 mg/kg and TD₅₀ (motorimpairment)=26.3 mg/kg.

Compound 120 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflex epilepsy (Frings mice)following i.p. administration with an ED₅₀=4.77 mg/kg and TD₅₀ (motorimpairment) between 20 and 30 mg/kg.

Compound 122 possessed the following additional biological activities:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflect epilepsy (Frings mice)following i.p. administration with an ED₅₀=4.7 mg/kg and TD₅₀ (motorimpairment)=15.3 mg/kg.

Compound 138 possessed the following additional biological activities:significant anticonvulsant activity against maximal electroshock-inducedseizures in mice following i.p. administration with an ED₅₀=51.9 mg/kgand TD₅₀ (motor impairment)=100.7 mg/kg.

Compound 151 possessed the following additional biological activities:significant anticonvulsant activity against maximal electroshock-inducedseizures in mice following i.p. administration with an ED₅₀=36.5 mg/kgand TD₅₀ (motor impairment)=108.4 mg/kg; a significant increase inseizure threshold as indexed by the i.v. Metrazol test in mice at thedoses of 36.5 and 108 mg/kg i.p.

Compound 156 possessed the following additional biological activates:significant anticonvulsant activity against sound-induced seizures in agenetically susceptible mouse model of reflect epilepsy (Frings mice)following i.p. administration with an ED₅₀=5.0 mg/kg and TD₅₀ (motorimpairment)=17.4 mg/kg.

Taken together, the results obtained with these simplified syntheticarylalkylamines suggest that such simplified molecules do not interactspecifically with the arylalkylamine binding site on receptor-operatedCa²⁺ channels as do Compounds 1, 2 and 3. Specifically, Compounds 19-215bind to the-site labeled by [³H]MK-801 at concentrations rangingapproximately 1 to 400-fold higher than those which antagonize thefunction of the NMDA receptor-ionophore complex. The fact that Compounds19-215 at therapeutic doses do not generally produce PCP-likestereotypic behavior, substitute for PCP in drug discrimination assays,or elicit neuronal vacuolization suggests, however, that such compoundsmight be useful either as lead compounds or drug candidates forneurological disorders and diseases. It has been reported that compoundswhich bind with low affinity (relative to MK-801) to the site labeled by[³H]MK-801 might possess therapeutic utility and possess a morefavorable side effect profile than that possessed by a high affinityantagonist such as MK-801 itself (Rogawski, Therapeutic potential ofexcitatory amino acid antagonists: channel blockers and2,3-benzo-diazepines. Trends Pharmacol. Sci. 14: 325, 1993). The lowaffinity of certain compounds within the group of Compounds 19-215(relative to MK-801) for the site labeled by [³H]MK-801 may place thesecompounds into this general class of low affinity noncompetitiveantagonists.

Identification of a Novel Modulatory Site on Receptor-Operated CalciumChannels

Having identified arylalkylamines which have therapeutically usefulproperties as defined above, compounds can now be identified which actat the critical arylalkylamine binding site on receptor-operated Ca²⁺channels, such as those present within NMDA, AMPA and nicotiniccholinergic receptor-ionophore complexes.

Examples of suitable tests now follow:

EXAMPLE 24 Radioligand Binding in Rat Cortex or Cerebellum

The following assay can be utilized as a high throughput assay to screenproduct libraries (e.g., natural product libraries and compound files atmajor pharmaceutical companies) to identify new classes of compoundswith activity at this unique arylalkylamine site. These new classes ofcompounds are then utilized as chemical lead structures for a drugdevelopment program targeting the arylalkylamine binding site onreceptor-operated Ca²⁺ channels. The compounds identified by this assayoffer a novel therapeutic approach to treatment of neurologicaldisorders or diseases. Examples of such compounds include those providedin the generic chemical formulae above. Routine experiments can beperformed to identify those compounds having the desired activities.

Rat brain membranes are prepared according to the method of Williams etal. (Effects of polyamines on the binding of [³H]MK-801 to the NMDAreceptor: Pharmacological evidence for the existence of a polyaminerecognition site. Molec. Pharmacol. 36: 575, 1989) with the followingalterations: Male Sprague-Dawley rats (Harlan Laboratories) weighing100-200 g are sacrificed by decapitation. The cortex or cerebellum from20 rats are cleaned and dissected. The resulting brain tissue ishomogenized at 4° C. with a polytron homogenizer at the lowest settingin 300 ml 0.32 M sucrose containing 5 μM K-EDTA (pH 7.0). The homogenateis centrifuged for 10 min at 1,000×g and the supernatant removed andcentrifuged at 30,000×g for 30 minutes. The resulting pellet isresuspended in 250 ml 5 μM K-EDTA (pH 7.0) stirred on ice for 15 min,and then centrifuged at 30,000×g for 30 minutes. The pellet isresuspended in 300 ml 5 μM K-EDTA (pH 7.0) and incubated at 32° C. for30 min. The suspension is then centrifuged at 100,000×g for 30 min.Membranes are washed by resuspension in 500 ml 5 μM K-EDTA (pH 7.0),incubated at 32° C. for 30 min, and centrifuged at 100,000×g for 30minutes. The wash procedure, including the 30 min incubation, isrepeated. The final pellet is resuspended in 60 ml 5 μM K-EDTA (pH 7.0)and stored in aliquots at −80° C. The extensive washing procedureutilized in this assay was designed in an effort to minimize theconcentrations of glutamate and glycine (co-agonists at the NMDAreceptor-ionophore complex) present in the membrane preparation.

To perform a binding assay with [³H]arylalkylamine, aliquots of SPMs(Synaptic Plasma Membranes) are thawed, resuspended in 30 mls of 30 μMEPPS/1 mM K-EDTA, pH 7.0, and centrifuged at 100,000×g for 30 minutes.SPMs are resuspended in buffer A (30 μM EPPS/1 μM K-EDTA, pH 7.0). The[³H]larylalkylamine is added to this reaction mixture. Binding assaysare carried out in polypropylene test tubes. The final incubation volumeis 500 μl. Nonspecific binding is determined in the presence of 100 μMnonradioactive arylalkylamine. Duplicate samples are incubated at 0° C.for 1 hour. Assays are terminated by the addition of 3 ml of ice-coldbuffer A, followed by filtration over glass-fiber filters (Schleicher &Schuell No. 30) that are presoaked in 0.33% polyethyleneimine (PEI). Thefilters are washed with another 3×3 ml of buffer A, and radioactivity isdetermined by scintillation counting at an efficiency of 35-40% for ³H.

In order to validate the above assay, the following experiments are alsoperformed:

(a) The amount of nonspecific binding of the [³H]arylalkylamine to thefilters is determined by passing 500 μl of buffer A containing variousconcentrations of [³H]larylalkylamine through the 10 presoakedglass-fiber filters. The filters are washed with another 4×3 ml ofbuffer A, and radioactivity bound to the filters is determined byscintillation counting at an efficiency of 35-40% for ³H. In filtersthat are not pretreated with 0.33% PEI, it was found that 87% of the³H-ligand was bound to the filter. Presoaking with 0.33% PEI reduces thenonspecific binding to 0.5-1.0% of the total ligand added.

(b) A saturation curve is constructed by resuspending SPMs in buffer A.The assay buffer (500 μl) contains 60 μg of protein. Concentrations of[³H]larylalkylamine are used, ranging from 1.0 nM to 400 μM in half-logunits. A saturation curve is constructed from the data, and an apparentK_(D) value and B_(max) value determined by Scatchard analysis(Scatchard, The attractions of proteins for small molecules and ions.Ann. N.Y. Acad. Sci. 51: 660, 1949). The cooperativity of binding of the[³H]arylalkylamine is determined by the construction of a Hill plot(Hill, A new mathematical treatment of changes of ionic concentrationsin muscle and nerve under the action of electric currents, with a theoryto their mode of excitation. J. Physiol. 40: 190, 1910).

(c) The dependence of binding on protein (receptor) concentration isdetermined by resuspending SPMs in buffer A. The assay buffer (500 μl)contains a concentration of [³H]arylalkylamine equal to its K_(D) valueand increasing concentrations of protein. The specific binding of[³H]arylalkylamine should be linearly related to the amount of protein(receptor) present.

(d) The time course of ligand-receptor binding is determined byresuspending SPMs in buffer A. The assay buffer (500 μl) contains aconcentration of [³H]arylalkylamine equal to its K_(D) value and 100 μgof protein. Duplicate samples are incubated at 0° C. for varying lengthsof time; the time at which equilibrium is reached is determined, andthis time point is routinely used in all subsequent assays.

(e) The pharmacology of the binding site can be analyzed by competitionexperiments. In such experiments, the concentration of[³H]arylalkylamine and the amount of protein are kept constant, whilethe concentration of test (competing) drug is varied. This assay allowsfor the determination of an IC₅₀ and an apparent K_(D) for the competingdrug (Cheng and Prusoff, Relationship between the inhibition constant(K_(i)) and the concentration of inhibitor which causes 50 percentinhibition (IC₅₀) of an enzymatic reaction. J. Biochem. Pharmacol. 22:3099, 1973). The cooperativity of binding of the competing drug isdetermined by Hill plot analysis.

Specific binding Of the [³H]arylalkylamine represents, binding to anovel site on receptor-operated Ca²⁺ channels such as those presentwithin NMDA-, AMPA and nicotinic cholinergic receptor-ionophorecomplexes. As such, other arylalkylamines should compete with thebinding of [³H]arylalkylamine in a competitive fashion, and theirpotencies in this assay should correlate with their inhibitory potenciesin a functional assay of receptor-operated Ca²⁺ channel antagonism(e.g., inhibition of NMDA receptor-induced increases in [Ca²⁺]_(I) incultures of rat cerebellar granule cells). Conversely, compounds whichhave activity at the other known sites on receptor-operated Ca 2,channels (e.g., MK-801, Mg²⁺, polyamines) should not displace[³H]arylalkylamine binding in a competitive manner. Rather, complexallosteric modulation of [³H]arylalkylamine binding, indicative ofnoncompetitive interactions, might be expected to occur. In preliminaryexperiments, MK-801 did not displace [³H]arylalkylamine binding atconcentrations up to 100 μM.

(f) Studies to estimate the dissociation kinetics are performed bymeasuring the binding of [³H]arylalkylamine after it is allowed to cometo equilibrium (see (d) above), and a large excess of nonradioactivecompeting drug is added to the reaction mixture. Binding of the[³H]arylalkylamine is then assayed at various time intervals. With thisassay, the association and dissociation rates of binding of the[³H]arylalkylamine are determined (Titeler, Multiple Dopamine Receptors:Receptor Binding Studies in Dopamine Pharmacology. Marcel Dekker, Inc.,New York, 1983). Additional experiments involve varying the reactiontemperature (0° C. to 37° C.) in order to understand the temperaturedependence of this parameter.

EXAMPLE 25 Radioligand Binding in Cerebellar Granule Cells

Primary cultures of cerebellar granule neurons are obtained from8-day-old rats and plated onto squares of Aclar plastic coated withpoly-L-lysine. The plastic squares are placed in 24-well culture plates,and approximately 7.5×10⁵ granule cells are added to each well. Culturesare maintained in Eagles' medium (HyClone Laboratories) containing 25 μMKCl, 10% fetal calf serum (HyClone Laboratories), 2 μM glutamine, 100μg/ml gentamicin, 50 U/ml penicillin, and 50 μg/ml streptomycin at 37°C. in a humid atmosphere of 5% CO₂ in air for 24 hr before the additionof cytosine arabinoside (10 μM, final). No changes of culture medium aremade until the cells are used for receptor binding studies 6-8 daysafter plating.

To perform a binding assay with [³H]arylalkylamine, the reaction mixtureconsists of 200 μl of buffer A (20 μM K-HEPES, 1 μM K-EDTA, pH 7.0) ineach well of the 24-well plate. The [³H]arylalkylamine is added to thisreaction mixture. Nonspecific binding is determined in the presence of100 μM nonradioactive arylalkylamine. Triplicate samples are incubatedat 0° C. for 1 hour. Assays are terminated by manually scraping thecells off the Aclar squares and placing them into polypropylene testtubes. The membranes prepared from whole cells in this manner aresuspended in 10 ml of ice-cold buffer A, and filtered over glass-fiberfilters (Schleicher & Schuell No. 30) that are presoaked in 0.33% PEI.The filtersare washed with another 3×3 ml of buffer A, and radioactivityon the filters is determined by scintillation counting at an efficiencyof 35-40% for ³H. The assay may be terminated by centrifugation ratherthan filtration in order to minimize nonspecific binding.

Specific experiments to characterize and validate the assay areperformed essentially as above, except that cells are used in place ofmembranes for the initial binding. The binding assay allows for thedetermination of an IC₅₀ value and an apparent K_(D). for the competingdrug as described by Scatchard analysis (The attractions of proteins forsmall molecules and ions. Ann. N.Y. Acad. Sci. 51: 660, 1949).Cooperativity of binding of the competing drug is determined by Hillplot analysis (A new mathematical treatment of changes of ionicconcentrations in muscle and nerve under the action of electriccurrents, with a theory to their mode of excitation. J. Physiol. 40:190, 1910). The specific binding of the [³H]arylalkylamine representsbinding to a novel site on receptor-operated calcium channels.

EXAMPLE 26 Recombinant Receptor Binding Assay

The following is one example of a rapid screening assay for usefulcompounds of this invention. In this assay, a cDNA or gene cloneencoding the arylalkylamine binding site (receptor) from a suitableorganism such as a human is obtained using standard procedures. Distinctfragments of the clone are expressed in an appropriate expression vectorto produce the smallest polypeptide(s) obtainable from the receptorwhich retain the ability to bind Compound 1, Compound 2 or Compound 3.In this way, the polypeptide(s) which includes the novel arylalkylaminereceptor for these compounds can be identified. Such experiments can befacilitated by utilizing a stably transfected mammalian cell line (e.g.,HEK 293 cells) expressing the arylalkylamine receptor.

Alternatively, the arylalkylamine receptor can be chemically reactedwith chemically modified Compound 1, Compound 2 or Compound 3 in such away that amino acid residues of the arylalkylamine receptor whichcontact (or are adjacent to) the selected compound are modified andthereby identifiable. The fragments of the arylalkylamine receptorcontaining those amino acids which are determined to interact withCompound 1, Compound 2 or Compound 3 and are sufficient for binding tosaid molecules, can then be recombinantly expressed, as described above,using a standard expression vector(s).

The recombinant polypeptide(s) having the desired binding properties canbe bound to a solid phase support using standard chemical procedures.This solid phase, or affinity matrix, may then be contacted withCompound 1, Compound 2 or Compound 3 to demonstrate that those compoundscan bind to the column, and to identify conditions by which thecompounds may be removed from the solid phase. This procedure may thenbe repeated using a large library of compounds to determine thosecompounds which are able to bind to the affinity matrix, and then can bereleased in a manner similar to Compound 1, Compound 2 or Compound 3.However, alternative binding and release conditions may be utilized inorder to obtain compounds capable of binding under conditions distinctfrom those used for arylalkylamine binding (e.g., conditions whichbetter mimic physiological conditions encountered especially inpathological states). Those compounds which do bind can thus be selectedfrom a very large collection of compounds present in a liquid medium orextract.

Once compounds able to bind to the arylalkylamine binding polypeptide(s)described above are identified, those compounds can then be readilytested in the various assays described above to-determine whether they,or simple derivatives thereof, are useful compounds for therapeutictreatment of neurological disorders and diseases described above.

In an alternate method, native arylalkylamine receptor can be bound to acolumn or other solid phase support. Those compounds which are notcompeted off by reagents which bind other sites on the receptor can thenbe identified. Such compounds define novel binding sites on thereceptor. Compounds which are competed off by other known compounds thusbind to known sites, or bind to novel sites which overlap known bindingsites. Regardless, such compounds may be structurally distinct fromknown compounds and thus may define novel chemical classes of agonistsor antagonist which may be useful as therapeutics. In summary, acompetition assay can be used to identify useful compounds of thisinvention.

EXAMPLE 27 Patch-Clamp Electrophysiology Assay

The following assay is performed for selected compounds identified inthe above-mentioned radioligand binding assays as interacting in ahighly potent and competitive fashion at the novel arylalkylaminebinding site on receptor-operated Ca²⁺ channels, such as those presentin NMDA-, AMPA- or nicotinic cholinergic receptor-ionophore complexes.This patch-clamp assay provides additional relevant data about the siteand mechanism of action of said previously selected compounds.Specifically, the following pharmacological and physiological propertiesof the compounds interacting at the arylalkylamine binding site aredetermined, utilizing the NMDA receptor-ionophore complex as an exampleof receptor-operated Ca²⁺ channels, potency and efficacy at blockingNMDA receptor-mediated ionic currents, the noncompetitive nature ofblock with respect to glutamate and glycine, use-dependence of action,voltage-dependence of action, both with respect to onset and reversal ofblocking, the kinetics of blocking and unblocking (reversal), andopen-channel mechanism of blocking. Such data confirm that the compoundsinteracting at the arylalkylamine binding site retain the uniquebiological profile of the arylalkylamines, and do not have their primaryactivity at the known sites on the NMDA receptor-ionophore complex(glutamate binding site, glycine binding site, MK-801 binding site, Mg²⁺binding site, Zn²⁺ binding site, sigma binding site, polyamine bindingsite).

Patch-clamp recordings of mammalian neurons (hippocampal, cortical,cerebellar granule cells) are carried out utilizing standard procedures(Donevan et al., Arcaine blacks N-methyl-D-aspartate receptor responsesby an open channel mechanism: whole-cell and single-channel recordingstudies in cultured hippocampal neurons. Molec. Pharmacol. 41: 727,1992; Rock and Macdonald, Spermine and related polyamines produce avoltage-dependent reduction of NMDA receptor single-channel conductance.Molec. Pharmacol. 42: 157, 1992).

Alternatively, patch-clamp experiments can be performed on Xenopusoocytes or on a stably transfected mammalian cell line (e.g., HEK 293cells) expressing specific subunits of receptor-operated Ca²⁺, channels.In this manner, for example, potency and efficacy at various glutamatereceptor subtypes (e.g., NMDAR1, NMDAR2A through NMDAR2D, GluR1 throughGluR4) can be determined. Further information regarding the site ofaction of the arylalkylamines on these glutamate receptor subtypes canbe obtained by using site-directed mutagenesis.

EXAMPLE 28 Synthesis of Arylalkylamines

Arylalkylamines such as Compound 1, Compound 2 and Compound 3 aresynthesized by standard procedures (Jasys et al., The total synthesis ofargiotoxins 636, 659 and 673. Tetrahedron Lett. 29: 6223, 1988; Nason etal., Synthesis of neurotoxic Nephila spider venoms: NSTX-3 and JSTX-3.Tetrahedron Lett. 30: 2337, 1989). Specific examples of syntheses ofarylalkylamine analogs 4-18 are provided in co-pending application U.S.Ser. No. 08/485,038, filed Jun. 7, 1995, and co-pending InternationalPatent Application No. PCT/US94/12293, published as WO95/21612, filedOct. 26, 1994, hereby incorporated by reference herein in theirentirety.

EXAMPLE 29 Synthesis of Simplified Arylalkylamines

Synthesis of Compound 20 was accomplished as follows.

A solution of sodium hydride (1.21.g, 50 mmol) in dimethoxyethane wastreated with diethyl cyanomethyl-phosphonate (8.86 g, 50 mmol) and thereaction stirred 4 hr at room temperature. To this was added3,3′-difluoroben-zophenone (10 g, 46 mmol) in DME. The reaction wasstirred 24 hr at room temperature, quenched with H₂O, and partitionedbetween diethyl ether and water. The ether fraction was dried overNa₂SO₄, and concentrated. GC/MS of this material showed 90% of theproduct A and 10% starting benzophenone.

A solution of this material in ethanol with a catalytic amount ofPd(OH)₂ was hydrogenated at 55 psi hydrogen for 4 hr at roomtemperature. The reaction was filtered and the catalyst washed withethanol (3×). The filtrate and ethanol washes were combined andconcentrated. GC/MS of this material showed 90% of the product B and 10%of the starting benzophenone.

A solution of this material in THF was treated with 70 ml 1 M B₂H₆ (70mmol) in THF and refluxed 1 hr. After cooling, the reaction was treatedwith 6 N HCl (50 ml) and refluxed an additional hour. After cooling, thereaction was basified to pH 14 with 10 N NaOH and equilibrated withether. The ether layer was removed and washed with 10% HCl (3×). Theacidic washes were combined, basified to pH 14 with 10 N NaOH andextracted with dichloromethane (3×). The organic washes were combined,dried over Na₂SO₄, and concentrated to yield an oil. GC/MS of thismaterial showed 100% Compound 20. GC/EI-MS (R_(t)=7.11 min) m/z(relative intensity) 247 (M⁺, 31), 230 (100), 215 (30), 201 (52), 183(63), 134 (23), 121 (16), 101 (21), 95 (15), 77 (15) This material indiethyl ether was filtered and treated with 35 ml 1 M HCl in ether. Theprecipitate was collected, dried, and recrystallized from water-ethanolto afford 1.045 g of Compound 20, as the hydrochloride salt. ¹H-NMR(CDCl₃) d 8.28 (3H, br s), 7.28-7.17 (2 H, m), 7.02-6.86 (6 H, m), 4.11(1H, t, J=8 Hz), 2.89 (2H, br t, J=8 Hz), 2.48 (2H, br t, J=7 Hz);¹³C-NMR (CDCl₃) d 164.6, 161.3, 144.8, 144.7, 130.4, 130.3, 123.3,123.2, 114.7, 114.5, 114.1, 113.8, 47.4, 38.4, 32.7.

Synthesis of Compound 21, Compound 33 and Compound 34 was accomplishedas follows.

A 100 ml round-bottomed flask equipped with stir bar, septa, and argonsource was charged with Compound 1 (2.43 g, 10 mmol) in 30 ml THF. Thesolution was cooled to −78° C. and treated dropwise with 11 ml lithiumbis(trimethylsilyl) amide (1M in THF) (11 mmol). The reaction wasstirred at −78° C. for 30 min and treated dropwise with excessiodomethane (3.1 ml, 50 mmol). The reaction was stirred 30 min at −58°C. GC/EI-MS analysis of an aliquot from the reaction showed consumptionof the starting nitrile 1. The reaction was quenched with water, dilutedwith diethyl ether and transferred to a separatory funnel. The etherlayer was washed with 10% HCl (3×), brine (1×), dried with anhydrousMgSO₄, and concentrated to a brown oil. This material was distilled(Kugelrohr, 100° C.) at reduced pressure to afford 1.5 g of a clear oil.GC/EI-MS of this material showed it to contain the desired product 2,(R_(t)=7.35 min) m/z (rel. int.) 257 (M⁺, 3), 203 (100), 183 (59), 170(5), 133 (4), 109 (3); ¹H-NMR (CDCl₃) d 7.4-6.9 (8H, m), 4.01 (1H, d,J=10 Hz), 3.38 (1H, dq, J=7, 10 Hz), 1.32 (3H, d, J=7 Hz); ¹³C-NMR(CDCl₃) d 19.4, 30.5, 54.2, 114.5, 114.6, 114.7, 114.9, 115.0, 115.3,123.3, 123.4, 123.6, 123.7, 130.5, 130.6, 131.7.

Product 3 was synthesized by the catalytic reduction of 2 using Raneynickel in 95:5 EtOH:aqueous sodium hydroxide (2 Eq.) under 60 psihydrogen. GC/EI-MS (R_(t=)7.25 min) m/z (rel. int.) 261 (M⁺, 20), 244(35), 229 (16), 215 (17), 201 (80), 183 (100), 133 (42), 115 (27), 109(47), 95 (20); ¹H-NMR (CDCl₃) d 7.3-6.8 (8H, m), 3.62 (1H, d, J=10 Hz),2.70 (1H, M), 2.40 (2H, m), 1.73 (2H, m), 0.91 (3H, d, J=7 Hz). Notethat product 3 in this reaction sequence corresponds to Compound 21.

Product 2 in 10% IPA-hexane (100 mg/ml) was chromatographed, in 500 μMaliquots, through Chiral Cel OD (2.0×25 cm) using 10% IPA-hexane at 10ml/min measuring optical density at 254 nm. This afforded the twooptically pure enantiomers 4 and 5 (as determined by analytical chiralHPLC; Note, the stereochemistry of these two compounds has not beenassigned at this time). These two compounds were identical in theirGC/EI-MS and ¹H-NMR spectra as product 2 (data above).

Each of the enantiomers 4 and 5 was reduced separately using dimethylsulfideborane complex in the following manner. A solution of compound (4or 5) in THF was heated to reflux and treated with excess (2 Eq.) 1M (inTHF) dimethyl sulfideborane complex and the reaction refluxed 30 min.After this time the reaction was cooled to 0° C. and treated with 6 NHCl. The reaction was set to reflux 30 min. After this time the reactionwas transferred to a separatory funnel, basified to pH>12 with 1ON NaOH,and the product (6 or 7) extracted into ether. The ether layer waswashed with brine, dried over anhydrous MgSO₄ and concentrated to anoil. The product was purified by prep-TLC using 5% methanol-chlorform.Each of the individual enantiomers (6 and 7) were found to be identicalin their GC/EI-MS and ¹H-NMR spectra as product 3 (data above). Notethat products 6 and 7 in this scheme correspond to Compounds 33 and 34.Compound 33 HCl: mp=260-270° C. (dec), [^(oc)]₃₆₅26=+6.6 (c 1.0 inEtOH), [oc]_(D)26=+0.4 (c 1.0 in EtOH). Compound 34.HCl:[^(oc)]₃₆₅23=−6.1 (c 1.0 in EtOH), [^(oc)]_(D)23=+0.1 (c 1.0 in EtOH).Compound 33 HI: The free base of Compound 33 was dissolved in EtOH and47% hydriodic acid (1.1 equivt.) was added. The solvent was evaporatedunder vacuum and the resulting solid hydroiodide was recrystallizedtwice from heptane/EtOAc by slow evaporation: mp=195-197° C. Theabsolute configuration of compound 33-HI was determined to be R bysingle-crystal (monoclinic colorless needle, 0.50×0.05×0.03 mm) X-raydiffraction analysis using a Siemens R3 m/V diffractometer (3887observed reflections).

Synthesis of Compound 22 was accomplished as described below. Compound23 was synthesized in a similar manner.

Triethyl phosphonoacetate (17.2 g, 76.8 mmol) was slowly added to asuspension of sodium hydride (3.07 g, 76.8 mmol) inN,N-dimethylformamide (350 ml). After 15 minutes3,3′-difluorobenzophenone (15.2 g, 69.8 mmol) was added to the solutionand stirred an additional 18 hr. The reaction mixture was quenched withwater and partitioned between water and ether. The combined organiclayers were washed with brine and dried over anhydrous magnesiumsulfate. The solvent was evaporated in vacuo to give 19.7 g of ethyl3,3-bis(3-fluorophenyl)acrylate as a yellow oil.

To a solution of ethyl 3,3-bis(3-fluorophenyl)-acrylate (19.7 g, 68.4mmol) in 200 ml of ethanol was added palladium hydroxide on carbon (3.5g). The mixture was shaken under 60 psi of hydrogen for 3 hours, thenfiltered and evaporated in vacuo to give 19.5 g of product A as acolorless oil.

The ethyl ester A (19.2 g) was hydrolyzed by stirring for 6 days with 50ml of 10 N sodium hydroxide. The reaction mixture was then diluted with50 ml of water and acidified to pH 0 with concentrated HCl. The aqueousmixture was extracted 3 times with ether and the ether extracts driedover magnesium sulfate and evaporated to give 3,3-bis(3-fluorophenyl)propionic acid as a white powder.

3,3-Bis(3-fluorophenyl)priopionic acid (13 g, 49.6 mmol) was dissolvedin 50 ml (685 mmol) of thionyl chloride and stirred overnight at roomtemperature. The excess thionyl chloride was removed in vacuo on arotary evaporator to give 13.7 g of product B as a yellow oil.

To acid chloride B (13.7 g, 49 mmol) dissolved in 100 ml of dry THF wasadded iron(III) acetylacetonate (0.52 g, 1.47 mmol). Methylmagnesiumchloride (16.3 ml, 49 mmol) was then added over a period of 1 hr bysyringe pump. The reaction was stirred for an additional hour, thenquenched by pouring into ether/5% HCl. The ether layer was separated,washed with 5% HCl and saturated NaCl, and dried over sodium sulfate.The solvent was evaporated in vacuo to give4,4-bis(3-fluorophenyl)-2-butanone as a yellow oil. The crude oil waspurified on silica gel using heptane/ethyl acetate as the elutant.

To 4,4-bis(3-fluorophenyl)-2-butanone (5.7 g, 21.9 mmol) in 25 ml ofethanol was added pyridine (1.91 g, 24.1 mmol) and methoxylaminehydrochloride (2.01 g, 24.1 mmol). The reaction was stirred overnight atroom temperature, then poured into ether/5% HCl. The ether layer wasseparated, washed with 5% HCl and saturated NaCl, and dried over sodiumsulfate. The solvent was evaporated in vacuo to give 6.26 g of the0-methyl oxime of 4,4-bis(3-fluorophenyl)-2-butanone. To sodiumborohydride (4.1 g, 108.3 mmol) in 15 ml of THF was slowly addedzirconium tetrachloride (6.31 g, 27.1 mmol). This mixture was stirredfor 15 min, then the oxime (6.26 g, 21.7 mmol) in 6 ml of THF was addedover 5 min. After 3 hours of stirring at room temperature, the reactionwas worked up by slowly adding 50 μM sodium hydroxide followed by ether.The aqueous layer was extracted 4 times with ether, and the combinedether extracts were dried over sodium sulfate. The solvent wasevaporated in vacuo to give 5.3 g of Compound 22.

Synthesis of Compound 24 was accomplished as described below. Compounds25-29, 52-53, 65, 76-78, 83, 90, 96-9,7, 115, and 135-136 were preparedin a similar manner.

A suspension of magnesium turnings (0.95 g, 39.2 mmol) in 150 mlanhydrous diethyl ether was treated with 1-bromo-3-fluorobenzene 6.83 g,39.2 mmol) dropwise via syringe. After 1.5 hr the solution wastransferred via cannula to a flask containing o-anisaldehyde (5.0 g,36.7 mmol) in 100 ml anhydrous diethyl ether at 0° C. and stirred 2 hr.The reaction mixture was quenched with water and partitioned betweenwater and ether. The combined organic layers were washed with brine anddried over anhydrous magnesium sulfate to afford 7.90 g (93% yield) ofproduct A.

Pyridinium dichromate (16.0 g, 42.5 mmol) was added to a solution of thealcohol A (7.90 g, 34.0 mmol) in dichloromethane (100 ml), and thereaction was stirred 12 hr. Diethyl ether (300 ml) was added to thereaction mixture and the black solution was filtered through a silicagel plug (30 cm) and washed with an additional 500 ml ether. Afterevaporation of the solvent in vacuo, the solid was recrystallized fromacetone to give 7.45 g (95% yield) of product B.

Diethyl cyanomethylphosphonate (7.0 g, 39.5 mmol) was slowly added to asuspension of sodium hydride (1.58 g, 39.5 mmol) in 100 mlN,N-dimethylformamide. After 30 minutes the ketone 3 was added to thesolution and stirred an additional 2 hr. The reaction mixture wasquenched with water, and partitioned between water and ether. Thecombined organic layers were washed with brine and dried over anhydrousmagnesium sulfate. The solvent was evaporated in vacuo to give a paleyellow oil.

In a glass bomb, the oil was dissolved in 100 ml ethanol and 20 ml 1ONNAOH. A catalytic amount of Raney Nickel suspended in water (ca. 15 molpercent) was added to the solution. The reaction mixture was shakenunder 60 psi H₂ for 12 hr on a Parr Hydrogenator. After filtering offexcess Raney Nickel, the solution was extracted with chloroform. Thecombined organic layers were washed with brine and dried over anhydrousmagnesium sulfate. After filtration, the oil was run through a silicagel column in chloroform and methanol. The solvent was evaporated invacuo to give a pale yellow oil. GC/EI-MS (R,=8.10 min) m/z (rel.intensity) 259 (100), 242 (44), 213 (48), 183 (42), 136 (50), 109 (94),91 (60), 77 (25). The oil was then acidified with hydrogen chloride indiethyl ether. Evaporation of the ether afforded a pale yellow solidthat was recrystallized in hot acetonitrile to afford 3.45 g (42.1%yield) white needles of Compound 24, as the hydrochloride salt.

Compounds 101 and 103 were synthesized from Compounds 25 and 24,respectively, by cleavage of their 0-methyl ethers with boranetr4-bromide in the normal manner.

Synthesis of Compound 30 was accomplished as described below. Compound31 was prepared in a similar manner.

A suspension containing magnesium turnings (0.95 g, 39.1 mmol) in 150 mlanhydrous diethyl ether was treated with 1-bromo-3-fluorobenzene (6.85g, 39.1 mmol) dropwise via syringe. After 1.5 hr the solution wastransferred via cannula to a flask containing 3-chlorobenzaldehyde (5.0g, 35.6 mmol) in 100 ml anhydrous diethyl ether at 0° C. and stirred 2hr. The reaction mixture was quenched with water and partitioned betweenwater and ether. The combined organic layers were washed with brine anddried over anhydrous magnesium sulfate to afford 8.40 g (>99% yield) ofproduct A.

Pyridinium chlorochromate (15.0 g, 39.8 mmol) was added to a solution ofthe alcohol A (8.40 g, 35.5 mmol) in 100 ml dichloromethane and stirred18 hr. Diethyl ether (300 ml) was added to the reaction mixture and theblack solution was filtered through a silica gel plug (30 cm), andwashed with an additional 500 ml ether. After evaporation of the solventthe solid was recrystallized from acetone to give 6.31 g (76% yield) ofproduct B.

Diethyl cyanomethylphosphonate (5.2 g, 29.6 mmol) was slowly added to asuspension of sodium hydride (1.2 g, 29.6 mmol) in N,N-dimethylformamide(100 ml). After 30 minutes the ketone B was added to the solution andstirred an additional 6 hr. The reaction mixture was quenched with waterand partitioned between water and ether. The combined organic layerswere washed with brine and dried over anhydrous magnesium sulfate. Thesolvent was evaporated in vacuo to give a yellow oil.

In a glass bomb, the oil was dissolved in ethanol (100 ml) and 10N NaOH(20 ml). A catalytic amount of rhodium suspended on alumina (ca. 35 molpercent) was added to the solution. The reaction mixture was shakenunder 60 psi H₂ for 24 hr on a Parr Hydrogenator. After filtering offexcess catalyst, the solution was extracted with chloroform. Thecombined organic layers were washed with brine and dried over anhydrousmagnesium sulfate. After filtration and evaporation of the solvent invacuo, the oil was taken up in tetrahydrofuran (100 ml). Diborane (23.4ml, 1.0 M) was added and the solution was refluxed for 1.5 hr. Thesolvent was evaporated in vacuo and 6N HCl (50 ml) was added carefully.The solution was refluxed for 1 hr. After cooling, the mixture wasbasified with 10N NaOH to pH 14 and partitioned between dichloromethaneand water. The combined organic layers were dried over anhydrousmagnesium sulfate and filtered. After evaporation of the solvent, theyellow oil was run through a silica gel column in chloroform andmethanol. The solvent was evaporated in vacuo to give a yellow oil.GC/EI-MS (R_(t)=8.15 min) m/z (rel. intensity), 263 (17), 246 (21), 211(84), 196 (33), 183 (100), 165 (19), 133 (19). The oil was thenacidified with hydrogen chloride in diethyl ether. Evaporation of theether afforded 0.96 g of a white solid, Compound 30, as thehydrochloride salt.

Synthesis of Compound 35 was accomplished as described below. Compounds36-37 were prepared in a similar manner.

A solution of 3-fluorobenzaldehyde (3.0 g, 24.2 mmol) at 0° C. in 150 mldiethyl ether was treated with 3.0 M ethyl magnesium chloride (12.7 ml,25.4 mmol) in tetrahydofuran (THF) via syringe. After 4 hr, the reactionmixture was quenched with water and partitioned between water and ether.The combined organic layers were washed with brine and dried overanyhydrous magnesium sulfate to afford 4.25 g of product A.

Pyridinium chlorochromate (6.53 g, 30.3 mmol) was added to a solution ofA in dichloromethane (100 ml) and stirred 18 hr. Diethyl ether (300 ml)was added to the reaction mixture and the black solution was filteredthrough a silica gel plug (30 cm) and washed with an additional 500 mlether. After evaporation of the solvent the solid was recrystallizedfrom acetone to give 3.05 g of product B. The solvent was evaporated invacuo to give a pale yellow oil.

Diethyl cyanometliylphosphonate (4.7 g, 26.4 mmol) was slowly added to asuspension of sodium hydride (1.1 g, 26.4 mmol) in 100 mlN,N-dimethylformamide. After 30 minutes the ketone B was added to thesolution and stirred an additional 6 hr. The reaction mixture wasquenched with water and partitioned between water and ether. Thecombined organic layers were washed with brine and dried over anhydrousmagnesium sulfate. The solvent was evaporated in vacuo to give a yellowoil.

In a glass bomb, the oil was dissolved in 100 ml ethanol and 20 ml 1ONNaCH. A catalytic amount of Raney Nickel suspended in water (ca. 15 molpercent) was added to the solution. The reaction mixture was shakenunder 60 psi H₂ for 24 hr on a Parr Hydrogenator. After filtering offexcess catalyst, the solution was extracted with chloroform. Thecombined organic layers were washed with brine and dried over anhydrousmagnesium sulfate. After filtration, the oil was run through a silicagel column in chloroform and methanol. The solvent was evaporated invacuo to give a pale yellow oil. GC/EI-MS (R_(t)=3.45 min) m/z (rel.intensity) 167 (4), 150 (63), 135 (58), 109 (100), 96 (53), 75 (48). Theoil was then acidified with hydrogen chloride in diethyl ether.Evaporation of the ether left a pale yellow solid that wasrecrystallized in hot acetonitrile to afford 2.2 g of Compound 35, asthe hydrochloride salt.

Synthesis of Compound 38 was accomplished as described below.

To a solution of 3,3-bis(3-fluorophenyl)-propionitrile (1.5 g, 6.17mmol) in 250 ml of THF at −70° C. was added butyl lithium (4.25 ml inhexanes, 6.8 mmol) by syringe over 5 minutes. The solution was stirredfor 5 min then methyl iodide (1.75 g, 12.3 mmol) was added over 1 min.The reaction mixture was then allowed to warm up to room temperature andworked up by diluting with ether and washing with 5% HCl and water. Theether layer was dried over sodium sulfate and evaporated to give 1.5 gof time methylated nitrile as a yellow oil.

To the 3,3-bis(3-fluorophenyl) 2-methyl-propionitrile (1.46 g, 5.7 mmol)in 50 ml of dichloromethane at 0° C. was added diisobutylaluminumhydride (1.02 ml, 5.7 mmol) by syringe over a 10 min period. Thereaction was stirred for 30 min at 0° C. followed by 2 additional hoursat room temperature. The reaction was worked up by adding 200 ml of 10%HCl and stirring at 40° C. for 30 min followed by extraction of theproduct with dichloromethane. The organic layer was dried over sodiumsulfate and evaporated to give 1.36 g of the product A.

To a solution of the aldehyde A (1-36 g, 5.23 mmol) in 40 ml of ether at0° C. was added methylmagnesium bromide (5.23 ml in ether, 5.23 mmol).The reaction was stirred for 3 hr at room temperature, and then quenchedwith dilute HCl. The ether layer was separated, dried over sodiumsulfate and evaporated to give 1.48 g of4,4-bis(3-fluorophenyl)-3-methylbutan-2-ol.

To a solution of the alcohol (1.4 g, 5.07 mmol) in 300 ml ofdichloromethane was added pyridinium chlorochromate 1.2 g, 5.58 mmol),and the mixture was stirred overnight. The reaction was then dilutedwith 100 ml of ether and filtered through a silica plug. The solvent wasevaporated to give 1.39 g of product B.

The ketone B (1.3 g, 4.9 mmol) was added to a solution of methoxylaminehydrochloride (0.45 g, 5.3B mmol) and pyridine (0.44 mL, 5.38 mmol) in30 ml of ethanol, and stirred overnight. The ethanol was thenevaporated, and the residue taken up in ether and 10% HCl. The etherlayer was separated, washed once with 10% HCl, dried over sodium sulfateand evaporated to give 1.4 g of the 0-methyl oxime.

To a suspension of sodium borohydride (0.87 g, 23.1 mmol) in 5 ml of THFwas added zirconium tetrachloride (1.35 g, 5.8 mmol), and the solutionwas stirred for 15 min followed by the addition of another 5 ml of THF.The 0-methyl oxime (1.4 g, 4.6 mmol) in 5 ml of THF was then added, andthe mixture stirred overnight. The THF was removed by evaporation invacuo, and the residue treated with 10% sodium hydroxide. After thebubbling ceased ether was added and the layers separated. The aqueouslayer was extracted four times with ether, and the combined etherextracts were dried over sodium sulfate. The ether was evaporated togive 1.25 g of Compound 38.

Compound 32 and Compounds 39-53 were synthesized according to standardprocedures as described above.

Compounds 107, 116, 139, and 143 were prepared as syntheticintermediates used in the preparation of Compounds.32, 115, 20, and 25,respectively.

Compound 50 was also prepared using the chiral synthesis describedbelow.

To an ice-cold solution of N-benzyl-(S)-a-methylbenzylamine (18.0 g,85.2 mmol) in THF (75 ml) was added butyl lithium (2.5 M in hexane; 37.5ml, 93.8 mmol) via a syringe over a period of 10 min at such a rate asto keep the reaction temperature below 10° C. during the addition. Thereaction was then stirred at 0° C. for 15 min. The reaction was cooledto −78° C. in a dry ice/isopropanol bath and then a solution of benzylcrotonate (15.0 g, 85.2 mmol) in THF (100 ml) was added dropwise over aperiod of 45 min. The reaction was stirred at −78° C. for 15 min, andthen saturated NH₄Cl (50 ml) was added. The reaction mixture was thenquickly transferred to a separatory funnel containing saturated NaCl(500 ml) and ether (200 ml). The layers were separated and the aqueouslayer extracted with ether (200 ml). The combined organic layers weredried, evaporated, and chromatographed on silica gel (50 mm×30 cm)(hexane-ethyl acetate [20:1]) to yield 21.0 g, 63.7% of product A.¹H-NMR showed that the diastereoselectivity of the reaction is >90%.

A mixture of magnesium (2.58 g, 106 mmol), THF (200 ml), and1-bromo-3-fluorobenzene (18.60 g, 106.3 mmol) was refluxed for 45 min.While still under reflux, product A (16.45 g, 42.45 mmol) was added viasyringe with THF (25 ml) over a 2 min period. The reaction was refluxedfor 1 hr, and then allowed to cool to room temperature. Saturated NH₄Cl(aq., 200 ml) was added. The reaction mixture was then transferred to aseparatory funnel containing saturated NaCl(aq) (500 ml) and diethylether (200 ml). The layers were separated and the aqueous layerextracted with ether (200 ml). The combined organic layers were driedover sodium sulfate and evaporated to give 21.41 g of product B as ayellow liquid.

Product B (20.02 g, 42.45 mmol, theoretical) was dissolved in aceticacid (120 ml) and sulfuric acid (30 ml). The reaction was stirred at 90°C. for 1 hr. The acetic acid was rotary evaporated giving a brownsludge. This material was placed in an ice bath and cold water (400 ml)was added. The product immediately precipitated. To the reaction wasslowly added 10 N NaOH (150 ml) to neutral pH. Diethyl ether (200 ml)was added to this mixture. The mixture was shaken until there was noundissolved material. The ether layer was separated, washed with water2×100 ml), dried over sodium sulfate, and rotary evaporated yielding13.14 g (68.2% based on ester) of a thick brown oil. This oil was takenup in ether and converted to the hydrochloride salt with hydrogenchloride in diethyl ether to give product C as a yellow-white solid.

Product C (7.17 g, 14.6 mmol) was taken up in absolute ethanol (200 ml).Pearlman's catalyst (Pd(OH)₂/C; 2.00 g) was added. The reaction wasshaken under 70 psi hydrogen gas at 70° C. for 20 hr, and the reactionmixture was filtered through Celite. The filtrate was rotary evaporatedto give 3.54 g of a light yellow glass. This material was taken up indiethyl ether (100 ml) and was basified with 1 N NaOH (25 ml). The etherlayer was washed with water (1×25 ml), dried over sodium sulfate, androtary evaporated to give 2.45 g of a light yellow oil. This materialwas Kugelrohr distilled (90-100° C., 1 mm Hg) to give 1.17 g of acolorless liquid. This material was taken up in diethyl ether andconverted to the hydrochloride salt with ethereal hydrogen chloride.After rotary evaporation, the salt was recrystallized from 0.12 N HCl(200 mg/ml). The crystals were filtered off and were washed with cold0.12 N HCl yielding 0.77 g (18%) of Compound 50 as silvery whitecrystals (as the hydrochloride salt).

Compound 51 was synthesized in a similar manner to Compound 50 utilizingN-benzyl-(R)-α-methylbenzylamine as a chiral starting material.

Synthesis of Compound 54 was accomplished as described below.

To a solution of 3,3′-difluorobenzophenone (5 g, 22.9 mmol) and methylcyanoacetate (3.4 g, 34.4 mmol) in 15 ml of ether was added titaniumisopropoxide (16.9 ml, 57.25 mmol). This solution was stirred for 6 daysat room temperature then quenched with 0.5 mol of HCl in 300 ml ofwater. The mixture was diluted with 100 ml of ether, and the layersseparated. The ether layer was washed with 5% HCl and saturated brine,then dried over sodium sulfate. The solvents were evaporated in vacuo togive 8 g of product A.

Compound A was dissolved in 50 ml of isopropanol, followed by theaddition of a small amount of bromocresol green. Sodium cyanoborohydride(1.52 g, 24.2 mmol) was added all at once followed immediately with thedropwise addition of concentrated HCl, added at such a rate as to keepthe solution yellow. After hours the reaction was worked up bypartitioning between ether and water. The ether layer was washed withwater and saturated brine, dried over sodium sulfate, and concentratedto give the product B.

To a solution of lithium aluminum hydride (30.4 ml, 30.4 mmol) in THFwas added product B (1 g, 3.04 mmol) in 2 ml of THF over a period of 30seconds. This solution was stirred overnight at room temperature, thenquenched with the addition of 20 ml of ethyl acetate. The solvents werethen removed in vacuo, and the resulting oil was dissolved in aqueousHCl and acetonitrile. The product was then purified on a C-18 columnwith a gradient of 0.1% HCl to acetonitrile to give 82 mg of Compound54, as the hydrochloride salt. EI-MS m/z (relative intensity) 277 (M⁺,100), 260 (2.4), 242 (8.6), 229 (28), 215 (11.7), 204 (16), 183 (12),133 (9.5), 124 (14), 109 (6.8), 30 (22).

Compound 55 was synthesized analogously to Compound 21 except that ethyliodide was used in the alkylation step. GC/EI-MS (R_(t)=7.43 min) m/z(relative intensity) 275 (M⁺, 100), 258 (66), 229 (63), 204 (57) 201(72), 183 (84), 134 (57), 124 (68), 109 (98), 72 (72).

The synthesis of Compound 56 was accomplished as follows.

The alcohol A was synthesized from 3-fluorobromobenzene and3-fluoro-2′-methylbenzaldehyde as described for product A Ln thesynthesis of Compound 24.

The alcohol A (8.4 g, 36.2 mmol) was stirred with manganese dioxide(12.6 g, 144.8 mmol) in 100 ml of dichloromethane for 4 days. Thereaction mixture was then diluted with ether and filtered through a 0.2micron teflon membrane filter. The filtrate was concentrated to give 7.6g of the ketone B.

The substituted acrylonitrile C was synthesized as described for productA in the Compound 20 synthesis.

To the nitrile C (4 g, 15.7 mmol) in 240 ml of ethanol was added 2 g of10% palladium dihydroxide on carbon. This mixture was hydrogenated at60-40 psi for 3 days. The reaction mixture was then filtered andconcentrated. The resulting oil was dissolved in chloroform andchromatographed on silica gel (30% methanol/5% isopropylamine inchloroform) to give the amine. This amine was dissolved in aqueousHCl/acetonitrile and purified via HPLC on C-18 (10% acetonitrile/0.1%HCl to 50% acetonitrile/0.1% HCl over 60 min) then lyophilized to give800 mg of Compound 56, as the hydrochloride salt. GC/EI-MS. (R_(t)=7.39min) m/z (relative intensity) 261 (M⁺, 64), 244 (56), 229 (57), 215(100), 203 (53), 183 (21), 133 (39), 122 (31), 109 (32).

The synthesis of Compound 57 was accomplished as follows.

To a solution of 5-fluoro-2-methylbenzonitrile (5 g, 37 mmol) in 50 mlof THF was added 3-fluorophenylmagnesium bromide (46 ml, 40 mmol) andcopper (I) cyanide (0.27 g, 0.8 mmol) this solution was refluxed for 4hours, then poured into ether/20% HCl and stirred for a further 2 hours.The layers were separated, and the ether layer washed with water andsaturated brine. The solution was dried over sodium sulfate andconcentrated. The crude oil was purified on silica (hexane to 50%dichloromethane in hexane over 60 min) to give 6.7 g of the ketone A.

The ketone A was converted to Compound 57 as described for Compound 56.GC/EI-MS (R_(t)=7.35 min) m/z (relative intensity) 261 (M⁺, 52), 244,(41), 229 (67), 215 (100), 203 (42), 201 (42), 183 (21), 133 (45), 122(28), 109 (26).

The synthesis of Compound 58 was accomplished as follows.

To a solution of 5-fluoro-2-methylbenzoyl chloride (2.24 g, 13 mmol) in10 ml of dry THF was added iron III acetylacetonate (0.16 g, 0.44 mmol).The solution was cooled to 0° C., and a THF solution of5-fluoro-2-methylphenylmagnesium bromide (20 ml, 15.5 mmol) was added bysyringe over a period of 30 min. The reaction was stirred for another 30min, then poured slowly into ether/5% HCl. The ether layer wasseparated, washed with saturated brine, dried over sodium sulfate, andconcentrated to give 3.2 g of ketone A.

Dry THF (30 ml)-was cooled to −78° C. followed by the addition of butyllithium (5.85 ml, 14.6 mmol, 2.5 M solution in hexanes). Acetonitrile(0.76 ml, 14.62 mmol) was then added over a period of 2 min, thenallowed to stir at −78° C. for 15 min. To this solution was added ketoneA (3 g, 12.2 mmol) in 5 ml of THF. The solution was stirred for 30 minat −78° C. then allowed to warm to room temperature and stirredovernight. The reaction mixture was partitioned between ether and 5%HCl. The ether layer was separated, washed with saturated brine, driedover sodium sulfate, and concentrated to give 2.2 g of the nitrile B.

The nitrile B (1 g, 3.48 mmol) was dissolved in 30 ml of ethanol and 3ml of 10 N sodium hydroxide. To this solution was added 1 g of a 5%aqueous slurry of Raney nickel, and the mixture was hydrogenated at 60psi for 20 hours. The reaction was filtered and concentrated to a whitesolid. This residue was taken up in ether/water and the ether layerseparated. The ether solution was dried over sodium sulfate andconcentrated to give 0.96 g of the hydroxyamine C.

The hydroxyamine C (0.96 g, 3.3 mmol) was taken up in concentrated HCland heated to 70° C. which caused brief solution, and then precipitationof the alkene D. The alkene was collected by filtration and dissolved in30 ml of ethanol and 1 ml of conc. HCl. Palladium dihydroxide on carbon(0.4 g) was added to the solution, and the mixture hydrogenated at 60psi for hours. The product was isolated by filtering off the catalystand evaporating the solvent. The residue was dissolved in 0.1% HCl andacetonitrile, and purified on C-18 (15% acetonitrile/0.1% HCl toacetonitrile) to give 0.6 g of Compound 58, as the hydrochloride salt.GC/EI-MS (R_(t)=7.82 min) m/z (relative intensity) 275 (M⁺, 100), 258(20) 243 (74), 229 (38), 214 (65) 201 (31), 196 (32), 183 (20), 148(35), 138 (42), 133 (48), 122 (69), 109 (41).

Synthesis of Compound 59 was accomplished as follows.

Compound 20 (2.0 g, 7.05 mmol) was dissolved in abs. EtOH (200 ml) andcooled to 5-10° C. in an ice bath. Acetaldehyde (0.395 ml, 7.05 mmol,cooled to −4° C.) was added followed by nickel-aluminum alloy (200 mg,Fluka Chemika), and the reaction was hydrogenated on a Parr apparatus at50 psi for 2 hr. GC/MS showed 75% yield of the product and 2% of theN,N-diethyl side-reaction product. The reaction mixture was filteredthrough diatomaceous earth and the filtrate was evaporated under reducedpressure. The crude product was dissolved in isopropanol (5 ml)/ether(60 ml)/ethereal HCl (1 M), and then hexane (5 ml) was added to thecloud point. The cloudy mixture was filtered through paper, then hexane10 ml) was added to the cloud point, and the solution was filteredagain. The filtrate was stoppered and the product was allowed tocrystallize at room temperature. The crystals were collected and driedto provide 0.325 g (14.8% yield) of Compound 59, as the hydrochloridesalt (colorless needles).

The synthesis of Compound 60 was accomplished as follows. Compounds 66,69, 108, 123, 142, and 145 can be synthesized in a similar mannerstarting from Compounds 33, 50, 32, 60, 25 and 119, respectively.

Compound 20 (as the free base) (1.0 g, 4.0 mmol) was refluxed in ethylformate (150 ml) for 2 hr. The solvent was then removed under reducedpressure to provide 1.1 g, 99% yield of formamide A as a colorless oil.GC/MS showed the product to be 100.0% pure and was used in the followingstep without further purification.

The formamide A (1.1 g, 4.0 mmol) was dissolved in dry THF (100 ml) andheated to reflux (no condenser). Borane-methyl sulfide complex (1.2 ml,12 mmol, 10.5 M) was added dropwise over a period of 3 min to therefluxing solution. Reflux was maintained for approximately 15 min, opento the air, until the reaction volume was reduced to approximately 30ml. The reaction was then cooled in an ice bath, and ice (5 g, smallpieces) was carefully added followed by H₂O (25 ml) and conc. HCl (25ml). The acidic solution was refluxed for 30 min. The reaction mixturewas then cooled in an ice bath, basified with NaOH (1ON), extracted withether (3×100 ml), dried (Na₂SO₄, anhydrous) and evaporated under reducedpressure. The crude product was dissolved in ether (10 ml)/hexane (50ml) and ethereal HCL (1 M) was added dropwise to precipitate thehydrochloride salt. The salt was collected and recrystallized fromisopropanol (3 ml)/ether (40 ml) to provide 0.5 g of Compound 60, as thehydrochloride salt.

Alternatively, Compound 60 was synthesized from commercially availablestarting materials in the following four step reaction sequence. Thefirst intermediate in this synthetic route,ethyl-N-benzyl-N-methyl-3-aminopropionate, was prepared by conjugateaddition of N-benzylmethylamine to ethyl acrylate. The esterfunctionality of the first intermediate was then reacted with twoequivalents of Grignard reagent (prepared from 1-bromo-3-fluorobenzene)to provide N-benzyl-N-methyl-3-hydroxy-3(bis-3-fluorophenyl)propylamine. The Grignard reaction product was then dehydrated in amixture of 6N HCl/acetic acid to yieldN-benzyl-N-methyl-3-(bis-3-fluorophenyl)-2-propenamine. Catalytichydrogenation of this material as its hydrochloride salt in ethanol overPearlman's catalyst [Pd(OH₂)/C] provided, after, recrystallization fromethyl acetate, colorless, needles of Compound 60 as the hydrochloridesalt.

In a 500-mL, 3-necked flask equipped with thermometer, reflux condenser,and a 125-mL addition funnel [charged with ethyl acrylate (88.3 mL, 81.5g, 0.815 mol)] was placed N-benzylmethylamine (100 mL, 94.0 g, 0.776mol). The ethyl acrylate was added dropwise to the stirring reactionmixture over a period of 80 min. After stirring for 18 h at roomtemperature, the product was vacuum distilled and the fractioncontaining product was collected at 78-95° C. (0.12-0.25 mm Hg), (138 g,80% yield): Bp 78-95° C. (0.12-0.25 mm Hg); TLC, R_(£)0.23 [hexane-EtOAc(5:1)], R_(£)=0.57 [MeOH—CHCl₃ (100:5)] GC, t_(R)=6.06 min; MS, 221(M⁺), 206 (M-CH₃), 192 (M-C₂H₅), 176 (M-0C₂H₅), 144 (M-C₆H₅), 134[CH₂N(CH₃)CH₂Ph] 120 [N(CH₃)CH₂Ph], 91 (C₇H₇), 77 (C₆H₅), 42 (CH₂CH₂N);¹H NMR (free base, CDCl₃) d 1.25 ppm (t, J=7.1, 3H, CH₂CH ₃), 2.20 (s,3H, NCH ₃), 2.5I (t, J=7.3, 2H, COCH ₂), 2.74 (t, J=7.2, 2H, CH ₂N),3.51 (s, 2H, NCH ₂Ph), 4.13 (q, J=7.1_(i). 2H, OCH ₂CH₃), 7.18-7.35 (m,5H, ArH); ¹³C NMR (free base, CDCl₃) d 15.2 (CH₂ CH₃), 34.0 (COCH₂),42.9 (NCH₃), 53.8 (NCH₂), 61.4 (0CH₂CH₃), 63.1 (CH₂Ph), 128.0 (CH),129.2 (CH), 130.0 (CH), 139.9 (q), 173.7 (q).

In a 5-L, four-necked, round-bottom flask, under nitrogen, were placedMg [51.5 g, 2.12 mol, turnings, washed with THF (2×300 mL)] and THF (2L). An addition funnel was charged with 1-bromo-3-fluorobenzene (neat,392.8 g, 2.24 mol). One-twentieth of the bromide was added to themagnesium suspension followed by one crystal of iodine. After initiationof the Grignard reaction the remaining 1-bromo-3-fluorobenzene was thenadded to the refluxing mixture over a period of 50 min. The reaction wasrefluxed for an additional 45 min. To the refluxing solution of Grignardreagent was added a solution of ethylN-benzyl-N-methyl-3-aminopropionate (187.5 g, 0.847 mol) in THF (100 mL)over a period of 20 min. After the ester addition was complete, thereaction was refluxed for 1 h. The reaction was then cooled in an icebath. Saturated NH₄Cl (aq., 400 mL) and H₂O (400 mL) were added and themixture was transferred to a separatory funnel. The organic layer wasseparated and the aqueous layer was-extracted once with THF (400 mL).The combined organic layers-were washed with satd. NaCl (2×200 mL, aq.),dried (anh. Na₂SO₄), filtered through paper, and rotary evaporatedvacuum to yield 281.6 g (90%) of crude product as an orange, viscousoil. This material (281.6 g, 0.766 mol) was dissolved in acetonitrile(1.4 L). Concentrated hydrochloric acid (65.0 mL, 0.786 mol, 12N) wasadded to the stirring filtrate. The crystallizing mixture was thencooled to −20° C. for 17 h. The product was collected, washed with coldacetonitrile (800 mL), and dried to provide a white solid, 235.6 g (69%yield from the ester). For analytical purposes, the hydrochloride saltwas further purified by recrystallization from acetonitrile: mp 194-197°C. (uncorr.); TLC, R_(£)=0.23 (hexane-EtOAc (5:1)], R_(£)=0.85[MeOH—CHCl₃ (100:5)], R_(£)=0.72 [MeOH—CHCl₃ (100:3)]; GC, t_(R)=10.93min; MS, 367 (M⁺), 272 (M-C₆H₄F), 258 (M-CH₂Ph-H₂O), 219 [(C₆H₄F)₂CH],148 [CH₂CH₂N(CH₃)CH₂Ph], 134 [CH₂N(CH₃CH₂Ph], 91 (C₇H₇, 42 (CH₂CH₂N); ¹HNMR (free base, CDCl₃) d 2.18 (s, 3H, NCH ₃), 2.41 (m, 2H, CHCH ₂), 2.58(m, 2H, CH ₂N), 3.42 (s, 2H, CH ₂PH), 6.86 (dt, J₁=8.5, J₂=1.8, 2H,Ar—H), 7.18-7.30 (m, 10H, Ar—H), 8.33 (bs, 1H, OH); ¹³C NMR (free base,CDCl₃) d 35.6 (CHCH₂), 41.5 (CH₃, NCH₃), 54.3 (CH₂, CH₂N), 62.6(CH₂,CH₂Ph), 113.1 (d, J=23, CH, Ar—C_(5,5′)), 113.5 (d, J=23, CH),121.2 (d, J=3, CH), 127.5 (CH), 128.5 (CH), 129.2 (CH), 129.5 (CH),129.6 (CH), 137.0 (q), 150.2 (q), 162.8 (d, J=243, q, Ar—C_(3,3′)).

In a 5-L, 3-necked reaction vessel, equipped with an overhead mechanicalstirrer, reflux condenser, and thermometer, was placedN-benzyl-N-methyl-3-hydroxy-3-bis (3-fluorophenyl) propylaminehydrochloride (225.4 g, 0.559 mol), 6N HCl (1392 mL) and glacial HOAc(464 mL). The suspension was heated in a water bath (80-85° C.) andstirred for 18 h. After 18 h of heating, the reaction mixture was cooledin an ice/MeOH bath. Ethyl acetate (500 mL) was added to the cooledreaction mixture. NAOH (1ON, 1.7 L) was then added to the cooled mixtureover a period of 25 min at such a rate as to keep the temperature below40° C. The mixture was transferred to a 6-L separatory funnel. Theorganic layer was separated and the aqueous layer was extracted withethyl acetate (2×500 mL). The combined organic layers were washed withsatd. NaCl (2×100 mL, aq.), dried Na₂SO₄ (250 g), rotary evaporated, andthen dried under vacuum to provide 185.6 g (95% yield) of the free baseas a fluid, brownish-colored oil.

The material above was stirred with hexane (1.5 L). The resultingsolution was filtered through paper. 4M HCl in dioxane (146 mL) wasadded dropwise with stirring to the filtrate over a period of 5 min. Thesemi-translucent solvent was then decanted away from the light-yellowcolored, semisolid precipitate. The crude hydrochloride salt wasdissolved in refluxing ethyl acetate (600 mL) and was filtered. Thefiltrate was then thoroughly cooled in an ice bath, and hexane (110 mL)was slowly added, with vigorous stirring. After cooling in an ice bathfor 2 h, the entire flask filled with a white crystalline solid. Thismaterial was collected on a filter funnel, washed with ice-cold.hexane/ethyl acetate E(1:41, 400 mL], and dried to yield 128.7 g, 59.7%of a white solid. On standing the mother liquor precipitated another14.8 g of an off-white solid. Total yield 128.7 g+14.8 g=143.5 (67%). Mp141-142° C. (uncorr.); TLC, R_(£)=0.20 [hexane-EtOAc (5:1)], R_(f)=0.75[MeOH—CHCl₃ (100:5)], R_(f)=0.49 [MeOH—CHCl₃ (100:3)]; GC, t_(R)=10.40min; MS, 349 (M⁺), 330, 301, 281, 258 (M-CH₂Ph), 240, 229[M-N(CH₃)CH₂Ph], 201, 183, 146, 133, 109, 91 (CH₂C₆H₅), 65, 42(CH₂NHCH₃); ¹H NMR (free base, CDCl₃) d 2.20 ppm (s, 3H, NCH ₃), 3.08(d, J=6.8, 2H, CH ₂N), 3.47 (d, J<1, 2H, CH ₂Ph), 6.29 (t, J=6.8, 1H,CH), 6.85-7.04 (m, 6H, ArH), 7.19-7.35 (m, 7H, ArH).

N-Benzyl-N-methyl-3-bis(3-fluorophenyl)allylamine hydrochloride (120.0g, 0.311 mol) was dissolved in abs. EtOH (1250 mL). Pd(OH)₂/charcoal(10.0 g, ˜20% Pd. Fluka Chemical) was added. The reaction mixture wasstirred under a steady flow of hydrogen gas for 18 h at 25° C.(atmospheric pressure). The mixture was then filtered throughCelite®/fritted glass, the catalyst was washed with ETOH (2×50 mL), andthe solvent was removed under reduced pressure to yield 95.4 g, 103% ofcrude product. This material was dissolved in refluxing ethyl acetate(300 mL) with vigorous stirring and filtered. The flask was allowed tostand for 2 h at 25° C., during which time the hydrochloride salt beganto crystallize as needles. The flask was then cooled, the product wascollected, washed with ice-cold ethyl acetate (20 mL), and dried toyield 73.7 g, 80%, of Compound 60 as a white, crystalline solid. Mp129-130° C.; UV/Vis, e=2.1×10³ L mol⁻¹ cm⁻¹ (264 nm, EtOH, 25° C.,linear range: 0.05-0.20 mg/mL); TLC, R_(f)=0.00 [hexane-EtOAc (5:1)],R_(f)=0.07 [MeOH—CHCl₃ (100:5)], R_(f)=0.19 [MeOH—CHCl₃—NH₄0H(100:5:1)]; GC, t_(R)=7.45 min; MS, 261 (M⁺), 229, 215, 201, 183, 164,150, 138, 122, 101, 83, 75, 57, 42 [CH₂NHCH₃], ¹H NMR (HCl salt, CDCl₃+1gtt MeOD) δ 2.56 (m, 2H, NCH ₂), 2.60 (s, 3H, NCH ₃) 2.85 (t, J=8.0, 2H,CHCH ₂) 4.11 (t, J=8.0, 1H, CH), 6.87-6.98 (m, 4H, ArH), 7.06 (d, J=7.7,2H, Ar_(2,2,)H), 7.25 (dd, J₁=6, J₂=8, ArH); ¹³C NMR (HCl salt, CDCl₃+1gt MeOD) δ 30.9 (CH₂, CHCH₂), 32.7 (CH₃, NCH₃), 47.6 (CH, CHCH₂), 47.8(CH₂, CH₂N), 113.9 (J=21, ArC_(2,2′) or ArC_(4,4′)), 114.5 (d, J=22,ArC_(2,2′) or ArC_(4,4′)), 123.2 (d, J=3, Ar—C_(6,6′)), 130.3 (d, J=9,Ar—C_(5,5′)), 144.7 (d, J=7, Ar—C_(1,1′)), 162.9 (d, J=245,Ar—C_(3,3′)); IR: KBr pellet (cm⁻¹), 3436.9, 2963.4, 2778.5, 2453.7,1610.6, 1589.3, 1487.0, 1445.3, 1246.0, 764.5; solubility: 2 g/mL (H₂O),1 g/mL (ETOH); anal. calcd. for C₁₆H₁₇NF₂—HCl Karl Fischer: 0.26 H₂O):C, 64.37; H, 6.11; N, 4.69; found: C, 64.14; H, 6.13; N, 4.69.

Compound 105 was prepared by selective reduction of its correspondingalkene by catalytic hydrogenation over Pd/C.

Compound 61 was prepared from 2-bromo-4-fluoroanisole and3-fluorobenzaldehyde as described for Compound 24. GC/EI-MS (R_(t)=9.22min) m/z (relative intensity) 277 (M⁺, 74), 260. (46), 245 (35), 231(44), 229 (34), 217 (24), 203 (28), 201 (31), 183 (28), 154 (24), 133(19), 109 (100).

Compound 62 was prepared from 2-bromoanisole and 2-methoxybenzaldehydeas described for Compound 24. GC/EI-MS (R_(t)=9.30 min) m/z (relativeintensity) 271 (M⁺, 100), 254 (17), 240 (23), 225 (40), 223 (45), 207(22), 181 (32), 165 (31), 136 (48), 121 (98), 91 (83).

The synthesis of Compound 63 was accomplished as follows.

Alcohol A was obtained from 3-fluorobenzaldehyde as described forproduct A of the Compound 24 synthesis.

To alcohol A (10.275 g, 47 mmol) in 200 ml of ethanol was added 1.6 g of10% Pd/C and 1 ml of concentrated HCl. This mixture was hydrogenated for3 hr at 60 psi, then filtered and concentrated to give thediphenylmethane B.

Product-B (2.01 g, 9.86 mmol) was dissolved in 20 ml of THF and cooledto −78° C. Butyl lithium (4.4 ml, 10.8 mmol, 2.5 M in hexanes) was addedslowly by syringe, and then the reaction stirred for another 30 min at−78° C. To this orange solution was added cyclopentene oxide (0.9 ml,10.3 mmol). The reaction was allowed to stir 3 hours while warmingslowly to room temperature. The reaction was quenched with 150 ml of 10%HCl and extracted 3 times with ether. The ether layer was dried oversodium sulfate and concentrated to give 2.5 g of the alcohol C.

To the alcohol C (1 g, 3.5 mmol) in 10 ml of dry THF was addedtriphenylphosphine (1.37 g, 5.2 mmol) in 5 ml of THF and p-nitrobenzoicacid (0.87 g, 5.2 mmol) in 5 ml of THF. This solution was cooled to 0°C. followed by the addition of DEAD (0.82 ml, 5.2 mmol), and allowed tostir overnight. The reaction was partitioned between water and ether.The ether was removed in vacuo and the resulting oil was chromatographedon silica gel in hexane/ethyl acetate to yield 365 mg of the cis-ester.This ester was hydrolyzed in methanol with potassium carbonate bystirring overnight. After removal of the methanol, the residue was takenup in ether, washed with water, dried over sodium sulfate andconcentrated to give 250 mg of the cis alcohol D.

To the alcohol D (0.25 g, 0.9 mmol) in 5 ml of dry THF was addedtriphenylphosphine (342 mg, 1.3 mmol) in 5 ml of THF and phthalimide(191.3 mg, 1.3 mmol) in 5 ml of THF. This solution was cooled to 0° C.followed by the addition of DEAD (0.205 ml, 1.3 mmol), and allowed tostir overnight. The reaction was partitioned between water and ether.The ether was removed in vacua and the resulting oil was chromatographedon silica gel in hexane/ethyl acetate to yield 100 mg of the phthalimideE.

To a solution of the phthalimide E (100 mg) in 20 ml of ethanol wasadded 8.8 mg of hydrazine hydrate. The solution was refluxed for 5 hoursthen stirred at room temperature overnight. The reaction was worked upby adding 1 ml. of conc. HCl and filtering off the white solid. Theresulting solution was concentrated to dryness and the solid taken up inether and aqueous sodium hydroxide. The ether layer was dried oversodium sulfate and concentrated to a white solid. This was taken up in asmall amount of ether and treated with 10 drops of 1M HCl in ether.After stirring overnight, the white solid was collected by filtrationand dried to give 50 mg of Compound 63, as the hydrochloride salt.GC/EI-MS (R_(t)=9.22 min) m/z (relative intensity) 287 (M⁺, 45), 270(12), 201 (63), 183 (81), 133 (38), 109 (43), 83 (44), 56 (100), 43(37).

The synthesis of Compound 64 was done as described for Compound 63except that the inversion step (product C to D) was omitted in order toobtain the cis amine as the final product. GC/EI-MS (R_(t)=8.28 min) m/z(relative intensity) 287 (M⁺, 15), 270 (4), 201 (13), 183 (15),133.(11), 109 (16), 84 (43), 56 (100), 43 (32).

The synthesis of Compound 65 was accomplished as follows.

The ketone A was synthesized similarly to ketone B in the Compound 24synthesis using 2-methylphenylmagnesium bromide and 2-methylbenzaldehydeas starting materials. This ketone was converted to the final productusing the procedure outlined for Compound 58. GC/EI-MS (R_(t), =7.84min) m/z (relative intensity) 239 (M⁺, 88), 222 (14), 207 (100), 193(46), 178 (71), 165 (60), 130 (39), 120 (40), 115 (51), 104 (40),91(38), 77 (21).

Compound 119 was synthesized in a seven-step reaction sequence startingfrom commercially-available trans-3-fluorocinnamic acid. This syntheticroute is conceptually similar to that reported in the literature [U.S.Pat. No. 4,313,896 (1982)] for related analogs. However, the three finalsteps were performed using a significantly different reaction sequencethan that reported. The cinnamic acid was reduced and chlorinated inthree steps to the corresponding 3-(3-fluorophenyl)propylchloride. Thiscompound was brominated with NBS (N-bromosuccinimide) and the resultingtrihalide was then reacted with 3-fluorophenol. The resulting ether wasconverted to the final product using a Gabriel synthesis.

Trans-3-fluorocinnamic acid (25.0 g, 150.4 mmol) was dissolved in abs.EtOH (250 mL) and hydrogenated over 10% Pd/C (2.5 g) in a Parr apparatusat 60 psig, 50° C., for 1 h (hydrogen uptake: calcd 245 psig; found 260psig). The reaction mixture was filtered and evaporated to yield acrystalline product (23.0 g, 89%). GC, t_(R)=4.43 min; MS, 168 (M⁺).

Under a stream of dry nitrogen, at 0-10° C., a solution of3-fluorohydrocinnamic acid (22.0 g, 131 mmol) in THF (100 mL) was addeddropwise, over a period of 15 min, to a suspension of LiAlH₄, (4.23 g,111 mmol) in THF (200 mL). The reaction was heated to reflux for aperiod of 1 h and then worked-up according to Fieser & Fieser's Reagentsfor Organic Synthesis (Vol. 1, 1967) to provide a white solid (20.1 g,99%). GC, t_(R)=3.74 min; MS, 154 (M⁺).

A solution of 3-(3-fluorophenyl)-1-propanol (15.0 g, 97.4 mmol) andtriphenylphosphine (36.0 g, 137.3 mmol) in CCl₄, (150 mL) was refluxedfor 19 h. Additional P(C₆H₅)₃ (3×3.0 g, 3×11.4 mmol) was addedperiodically over a period of 24 h. The resulting precipitate wasremoved by filtration and the solids were washed with hexane. Thefiltrate was evaporated under vacuum and the residue was suspended inhexane (200 mL) and then filtered. Evaporation of the filtrate provided16.0 g (95.1%) of crude product which was purification by silica gelflash chromatography, elution with hexane, to provide 14.7 g (87%) of acolorless liquid. GC, t_(R)=3.63 min; MS, 172/174 (M⁺)

A solution of the above chloride (12.0 g, 69.5 mmol), N-bromosuccinimide(17.3 g, 97.2 mmol), and dibenzoyl peroxide (0.06 g) in CCl₄, (75 mL)was refluxed for 1 h. The reaction mixture was then cooled in an icebath, filtered, and the solids were washed with hexane. The filtrate wasevaporated to provide 17.9 g (100%) of product. GC, t_(R)=5.21 min; MS,251/253 (M⁺)

A mixture of 3-bromo-3-(3-fluorophenyl)-1-propylchloride (4.0 g, 15.9mmol), 3-fluorophenol (1.98g, 17.7 mmol), and K₂CO₃ (2.65 g, 19.2 mmol)suspended in acetone (80 mL) was refluxed for 15 h. The volatiles werethen removed under vacuum and the resulting residue was suspended in amixture of hexane (200 mL) and NaOH (0.1N, 100 mL). The layers wereseparated and the organic layer washed, 0.1N NaOH (100 mL) and H₂O (100mL), dried (anh. Na₂SO₄), and evaporated in vacuo. The resulting residuewas chromatographed on silica gel, elution with hexane followed byhexane/EtOAc [100:1] then [40:1] to provide 1.64 g (37%) of product as acolorless oil. GC, t_(R)=7.28 min; MS, 282/283 (M⁺); TLC r_(£)=0.3,hexane/EtOAc [40:1].

A solution of 3-(3-fluorophenyl)-3-(3-fluorophenoxy)-1-propylchloride(1.52 g, 5.38 mmol) and potassium phthalate (1.20 g, 6.48 mmol) washeated to 90° C. in DMF (30 mL) for a period of 2 h in a nitrogenatmosphere. The reaction mixture was then cooled and poured into H²O(100 mL). The resulting solution was extracted with Et₂O (2×100 mL). Theorganic extract was washed, sat. NaCl (100 mL) and H₂O (2×100 mL), dried(anh. Na₂SO₄), and evaporated under vacuum to provide 2.17 g of crudeproduct. The material was chromatographed on silica gel, elution withhexane/EtOAc (40:1] and then [20:1] to provide after evaporation 1.81 g(86%) of product as a glass.

A solution ofN-phthaloyl-3-(3-fluorophenyl)-3-(fluorophenoxy)-1-propylamine (1.74 g,4.42 mmol) and anh. hydrazine (1.43 g, 44.6 mmol) in abs. EtOH (30 mL)was refluxed for 1 h. The reaction was cooled and evaporated undervacuum. The resulting material was suspended in Et₂O (75 mL) and washedwith 0.2N NaOH (2×25 mL). The organic layer was dried (anh. Na₂SO₄), andevaporated under vacuum to provide 1.04 g (89.3%) which was purified byreverse-phase chromatography [Vydac Prep. C18; 264 nm; 50 mL/min;gradient elution ACN/0.1% HCl aq., 10%-50% over 20 min; r_(t)=17.4 min],to yield 0.89 g (67%) of Compound 119 as a hygroscopic hydrochloridesalt.

Compounds 118, 120-122 and 137 were prepared in a manner similar to theprocedures used for the preparation of Compound 119.

Compound 113 was synthesized from commercially available4,4-diphenylcyclohexenone in three steps. First, the alkene in thestarting material was reduced by means of catalytic hydrogenation.Methoxylamine formation followed by reduction using standard procedures.

The synthesis of Compounds 188 and 189 was accomplished as follows.

Compounds 188 and 189

The enantiomers of Compound 136 were separated by analytical chiralHPLC. Aliquots (20 μg) were injected onto a Chiralcel-OD-R (ChiralTechnologies, Inc., Exton, Pa.) reversed-phase HPLC column (0.46×250 mm)using the following conditions: gradient elution, 40%-70% ACN (60-30%0.5N KTFA) over 30 min; flow rate, 1 mL/min; detector, 264 nm. Twoidentically-sized peaks were collected at 21.0 and 24.4 min. GC/MSanalysis of the two samples indicate that both materials have identicalGC retention times as well as identical mass spectra.

The synthesis of Compound 151 was accomplished as follows.

3,3-Bis(3-fluorophenyl)propanamide (Compound 151)

A solution of liquid anh. ammonia (10 mL) in CH₂Cl₂ (50 mL) at −78° C.was treated with a solution of 3,3-bis(3-fluorophenyl)propionyl chloride(2.19 g, 7.81 mmol) in CH₂Cl₂ (25 mL). The reaction was then stirred atambient temperature for 15 min and was then diluted with diethyl ether(500 mL), washed three times with 10% HCl, three times with 1N NaOH, andfinally once with H₂O. The organic layer was dried (anh. Na₂SO₄) andevaporated to give the primary amide as a white solid (2.01 g, 98%).

The synthesis of Compound 156 was accomplished as follows.

5-Cyanomethylidino-10,11-dihydrodibenzo[a,d]cycloheptene

To a solution of diethyl cyanomethylphosphonate (9.66 g, 54.5 mmol) indry N,N-dimethylformamide (DMF, 40 mL) was added NaH (60% dispersion,2.20 g, 55.0 mmol) over a period of 2 min. The reaction was stirred for10 min and then a solution of dibenzosuberone (10.3 g, 49.6 mmol) in dryDMF (10 mL) was added over a period of 2 min. The reaction was stirredat 80° C. for 4 h under N₂. Water (200 mL) was added and the reactionmixture was extracted with Et₂O (2×100 mL). The combined organic layerswere rotary evaporated to less than 50 mL. The resulting crystals werecollected and washed with cold Et₂O (2×50 mL) to yield 7.48 g (65.3%).

5-(2-Aminoethyl)-5H-10,11-dihydrodibenzo[a,d]cycloheptene hydrochloride(Compound 156)

5-Cyanomethylidino-10,11-dihydrodibenzo[a,d]cycloheptene was dissolvedin EtOH (100 mL). 1N NaOH (10 mL) and Raney® nickel (aq. suspension,0.50 g) were added. The reaction mixture was shaken under 60 psig H² at50° C. for 22 h, and was then filtered through Celite®. The filtrate wasrotary evaporated and the residue was dissolved in Et₂O (100 mL), washedwith satd. aq. NaCl (50 mL) and H₂O (50 mL). The Et₂O layer was dried(anh. Na₂SO₄) and rotary evaporated to give the crude product (850 mg)as a colorless oil. This oil was dissolved in EtOAc (5 mL) and filtered.1.OM HCl (5 mL) in Et₂O was added to the filtrate and a white,crystalline solid precipitated. This material was recrystallized fromEtOH (5 mL)-Et₂0 (12 mL) to yield 600 mg (50.7%) of product as a whitepowder.

The synthesis of Compound 167 was accomplished as follows.

2-Methoxypropiophenone

A mixture of 2-hydroxypropiophenone (3.00 g, 20.0 mmol), iodomethane(3.40 g, 24.0 mmol), and K₂C0₃ (granular, anh.; 13.8 g, 99.9 mmol) wasrefluxed in acetone (75 mL) for 18 h. The reaction mixture was cooled toroom temperature and the inorganic salts were removed by filtration. Thefiltrate was evaporated under vacuum to give an oil which wassubsequently dissolved in diethyl ether (200 mL) and then washed with0.1N NaOH (3×50 mL) followed by H₂O (50 mL). The organic layer was dried(anh. Na₂SO₄), filtered, and evaporated to an orange oil (3.17 g,96.9%). This material was used in the following step without furtherpurification. TLC, R_(f) 0.55 (1% MeOH:1% IPA:CHCl₃) GC, tr 4.58 min;MS, m/z 164 (M+1).

(R,S)-N-1-(2-methoxyphenylpropyl)-3,3-diphenylpropylamine (Compound 167)

A solution of 2-methoxypropiophenone (0.848 g, 5.17 mmol),3,3-diphenylpropyl amine (1.00 g, 4.70 mmol), and titanium(IV)isopropoxide [Ti(OCH(CH₃)₂)₄; (1.76 mL, 5.88 mmol, 1.25 equiv)] wasstirred at room temperature for 6 h. EtOH (2 mL)) was then added,followed by sodium cyanoborohydride (0.295 g, 4.70 mmol) in portionsover a period of 10 min, and the reaction was then stirred for 18 h. Thereaction mixture was then poured into diethyl ether (200 mL) and theresulting suspension was centrifuged to remove the titanium precipitate.The supernatant was collected and the pellet was rinsed with diethylether (200 mL). The combined organic washings were evaporated undervacuum to give a crude oil which was chromatographed on silica gel(elution with 4% MeOH—CH₂Cl₂). to provide 647 mg (38%) of product. Thematerial was then dissolved in diethyl ether (50 mL), filtered, andexcess ethereal HCl was added to precipitate the hydrochloride salt (125mg, 7.4%) as a white solid; TLC, R_(f) 0.25 (4% MeOH—CH₂Cl₂); GC,t_(r)=11.2 min; MS, m/z 359 (M+) The synthesis of Compounds 172-176 wasaccomplished as follows.

R,S) -3,3′-Difluoro-4-methoxybenzhydrol

A mixture of Mg° turnings (2.45 g, 101 mmol), 1-bromo-3-fluorobenzene(17.69g, 100 mmol), and dry THF (200 mL) was carefully heated to refluxfor 30 min. While still refluxing, 3-fluoro-p-anisaldehyde (15.3 9, 99.3mmol) in THF (100 mL) was added over a period of 5 min. The reactiontemperature was maintained for 30 min. cooled to room temperature, andthen the reaction was quenched with satd. aq. NH₄Cl (200 mL). Theorganic layer was separated, washed with satd. aq. NaCl (2×200 mL),dried (anh. Na₂SO₄) and rotary evaporated to yield 23.5 g (94.4%) ofproduct as an orange-brown oil.

3,3 ′-Difluoro-4-methoxybenzophenone

Pyridinium chlorochromate (22.3 g, 103 mmol) was added to a solution of3,3′-difluoro-4-methoxybenzhydrol (23.5 g, 93.8 mmol) in CH₂—Cl₂ (300mL) and the reaction mixture was stirred for 16 h. Diethyl ether (500mL) was added and the reaction mixture was filtered through Celite. Thefiltrate was rotary evaporated and the resulting oil was flashchromatographed (gradient elution of hexanes to 1:1 hex-EtOAc). TheTLC-pure fractions were rotary evaporated to yield 1.58 g of a whitesolid. The rest of the impure fractions containing product were combinedand rotary evaporated to the point where crystals began to form.Additional hexane (300 mL) was added and the crystallizing solution wasallowed to stand. The resulting crystals were collected and washed withhexanes (2×50 mL) to yield 6.81 g of product. The two batches werecombined to afford a total yield of 8.39 g (36.1%).

(R,S)-α-Cyanomethyl-3,3′-difluoro-4-methoxybenzhydrol

To dry THF (100 mL) was added butyllithium (2.6M in heptane; 16.0 mL,41.6 mmol) at −78° C. Acetonitrile 30 (2.20 mL, 42.1 mmol) was addedover a period of 1 min and the reaction was stirred at −78° C. under N₂for 30 min. A solution of 3,3′-difluoro-4-methoxybenzophenone (8.38 g,33.8 mmol) in anh. THF (50 mL) was added to the reaction over a periodof 5 min and the solution was stirred at −78° C. for 30 min. The coldbath was removed and the reaction was allowed to warm for 30 min. Satd.aq. NH₄C₁ (100 mL) was added to quench the reaction. The THF layer wasseparated, washed with satd. aq. NaCl (2×25 mL), dried (anh. Na₂SO₄),rotary evaporated, and dried under vacuum to yield 10.1 g (103%) ofproduct as a yellow oil.

(E)- and (Z)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)allylaminehydrochloride (Compound 172)

(R,S)-α-Cyanomethyl-3,3′-difluoro-4-methoxybenzhydrol (9.77 g, 33.8mmol) was dissolved in dry THF (200 mL) and heated to boiling (nocondenser). Under a stream of nitrogen, borane-dimethyl sulfide complex(BH₃ S(CH₃) 2, 10.1M; 16.8 mL, 170 mmol) was added carefully over aperiod of 2 min to the boiling solution. Boiling was then maintained for15 min until most of the THF was gone. The reaction mixture was thencooled in an ice bath. Ice (10 g) was carefully added, followed by H₂O(50 mL). The reaction was then heated to near boiling and 12.1N HCl (100mL) was added. The reaction was boiled (no condenser) for 30 min and wasthen cooled in an ice bath, basified with 10N NaOH (100 mL) andextracted twice with Et₂O (200 mL, 100 mL). The combined ether layerswere washed with 1N NaOH (50 mL) and H₂O (50 mL) dried (anh. Na₂SO₄),and rotary evaporated. The resulting oil was flash chromatographed(CHCl₃; 1:100 MeOH—CHCl₃; 1:10 MeOH—CHCl₃) through flash silica gel toafford 6.83 g of a yellow oil. This oil was dissolved EtOH (2 mL) andEt₂O (10 ML). 1.0 HCl in Et₂O (27 ML) was added and the solution wasrotary evaporated to yield 7.10 g (67.5%) of product as a solid, yellowfoam.

(R,S)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)propylamine maleate(Compound 173)

The mixture of (E)- and(Z)-3-(3-fluoro-4-methoxy)-3-(3-fluorophenyl)-allylamine hydrochlorides(7.10 g, 22.8 mmol) was dissolved in ETOH (200 mL) and a suspension ofpalladium on charcoal (10% Pd; 0.71 g) in H²O (3.5 mL) was added. Thereaction mixture was then shaken under 60 psig H₂ for 18 h andsubsequently filtered through Celite. The filtrate was rotaryevaporated, the residue was dissolved in EtOAc (25 mL) and Et₂O (100mL), and was basified with sat. aq. NaHCO₃ (25 mL). The organic layerwas separated, dried (anh. Na₂SO₄), and rotary evaporated to yield 6.28g of an oil. This oil and maleic acid (2.59 g) were dissolved into hotEtOAc (100 mL). Diethyl ether (70 mL) was added and crystals soon beganto form. The crystals were collected and dried to yield 2.45 g (27.3%)of a white powder. The combined filtrate and washings afforded morecrystalline product out upon standing. The second crop was filtered,washed with 1:1 EtOAc-Et₂O (2×25 mL) and Et₂O (1×25 mL), and dried toprovide 3.69 g (41.2%) of a white powder. The total yield was thus 6.14g (68.5%).

(R,S)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)propylformamide

(R,S)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)propyl amine maleate(3.12 g, 7.93 mmol) was free-based in a mixture of EtOAc (25 mL), Et₂O(100 mL), and satd. aq. NaHCO₃ (25 mL). The organic layer was separated,dried (anh. Na₂SO₄), and rotary evaporated. A solution of the amine inethyl formate (75 mL, 930 mmol) was refluxed for 17 h. The reactionsolution was rotary evaporated to yield 2.38 g (98.3%) of formamide as alight-orange, viscous oil.

(R,S)-N-Methyl-3-(3-fluoro-4-methoxy)-3-(3-fluorophenyl)propyl aminemaleate (Compound 174)

(R,S)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)propyl formamide (2.27,7.43 mmol) in THF (100 mL) was heated to boiling (no condenser).Borane-dimethyl sulfide complex (10.1M; 2.30 mL, 23.2 mmol) was addedcarefully over a period of 2 min to the boiling solution. Boiling wasthen maintained for 15 min. The reaction was then cooled in an ice bath.Ice (10 g) was carefully added, followed by H₂O (30 mL), followed by12.1N HCl (50 mL). The reaction was then boiled (no condenser) for 30min. The reaction was subsequently cooled in an ice bath, basified with1ON NaOH (50 mL), and extracted with Et₂O (200 mL). The ether layer waswashed with satd. aq. NaCl (100. mL), dried (anh. Na₂SO₄), and rotaryevaporated to yield 2.03 g of a yellow oil. This material was purifiedby RP-HPLC (20-60% acetonitrile-0.1% aq. HCl over 20 min). The collectedfractions were frozen and lyophilized to yield 1.28 g of a white solid.The free-base of the purified amine was dissolved in EtOAc. Maleic acid(305 mg) was added and the mixture was heated until everything haddissolved. The product was crystallized by adding Et₂O (5 mL). Thecrystals were filtered and washed with 1:1 EtOAc-Et₂O (10 mL) followedby Et₂O (10 mL) to yield 967 mg of product as a white, finelycrystalline solid. (R,S)-3-(3-Fluoro-4-hydroxy)-3-(3-fluorophenyl)propylamine maleate (Compound 175)

(R,S)-3-(3-Fluoro-4-methoxy)-3-(3-fluorophenyl)propyl-amine maleate(2.45 g, 6.23 mmol) was free-based in the normal manner and dissolved inCH₂Cl₂ (25 mL) The resulting solution was cooled to −78° C. Under N₂flow, boron tribromide (1-OM in CH₂Cl₂; 15 ml, 15 mmol) was added over aperiod of 5 min. The cold bath was removed and the reaction mixture wasallowed to warm to room temperature. After 30 min at 25° C., thereaction was hydrolyzed with 12.1N HCl 10 mL). The aqueous layer wasneutralized (pH 7) by the careful addition of 1ON NaOH (14 mL). Satd.aq. NaHCO₃ (50 mL) was added along with Et₂O (100 mL) and EtOAc (20 mL).This mixture was shaken vigorously and the organic layer was separated.The aqueous layer was extracted with EtOAc (20 mL). The combined organiclayers were dried (anh. Na₂SO₄) and rotary evaporated. The resulting oilwas dissolved in EtOH, 1.OM HCl in Et₂O (7 mL) was added, and thesolution was rotary evaporated. This material was then purified byRP-HPLC [20-60% acetonitrile-0.1% HCl (aq.) over 20 min]. The collectedfractions were frozen and lyophilized, affording 716 mg of a whitesolid. The free-base of the purified amine (315 mg) was dissolved inEtOAc. Maleic acid (138 mg) was added and the mixture was heated untileverything dissolved. The EtOAc was rotary evaporated to give a hardglass which was dissolved in MeOH (5 mL). Water (100 mL) was then addedand the solution was subsequently frozen and lyophilized. The aboveprocedure yielded 445 mg of product as a white solid.

(R,S)-N-Methyl-3-(3-fluoro-4-hydroxy)-3-(3-fluorophenyl)propyl aminehydrochloride (Compound 176)

A solution of(R,S)-N-methyl-3-(3-fluoro-4-methoxy)-3-(3-fluorophenyl)propylamine (703mg, 2.41 mmol) in CH₂Cl₂ (10 mL) was cooled to −78° C. Under nitrogen,boron tribromide (1.OM in CH₂Cl₂; 6.0 mL, 6.0 mmol) was added over aperiod of 5 min. The cooling bath was then removed and the reaction wasallowed to warm to room temperature. After 1 h, the reaction wasquenched with 12.1N HCl (5 mL). The aqueous layer was then neutralized(pH 7) by the careful addition of 1ON NaOH (˜7 mL). Satd. aq. NaHCO₃ (25mL) was added along with Et₂O (50 mL), EtOAc (15 mL), and CHCl₃ (5 mL).This mixture was shaken vigorously, and the organic layer was separated,dried (anh. Na₂SO₄), and filtered through paper. The crude product wasthen purified by RP-HPLC (20-60% acetonitrile-0.1% aq. HCl gradient over20 min). The fractions were frozen and lyophilized to afford 602 mg ofproduct as a white solid.

The synthesis of Compound 185 was accomplished as follows.

(R)-3-(3-Fluorophenoxy)-3-phenylpropylchloride

Following a similar procedure for the chiral synthesis of fluoxetine[Srebnik, M. et al., J. Org. Chem. 25.53(13), 2916-20 (1988), herebyincorporated by reference herein], a solution of(S)-(−)3-chloro-1-phenyl-1-propanol (4.00 g, 23.4 mmol), 3-fluorophenol(2.63 g, 23.4 mmol), and diethyl azodicarboxylate (4.00 g, 23.4 mmol)were dissolved in THF (200 mL). The mixture was cooled to 0° C. andtriphenylphosphine (6.77 g, 25.8 mmol, 1.1 equiv) was added slowly over10 min. The reaction mixture was then stirred at room temperature for 18h. The THF was subsequently evaporated under vacuum to afford a gelwhich was washed with pentane (3×50 mL). The pentane washings werefiltered and the filtrate was evaporated under vacuum to give a clearoil. This oil was dissolved in diethyl ether (150 mL) and washed with 1%HCl-satd. NaCl (25 mL), 0.1N NaOH-satd. NaCl (2×25 mL), and finally H₂O(2×25 mL). The organic layer was then dried (anh. Na₂SO₄), filtered, andevaporated to dryness under vacuum to give an orange oil. The crudeproduct was chromatographed on silica gel (25×180 mm, gravity column),elution with 40:1 hexane-EtOAc, to provide 971 mg (15.7%) of product asa colorless oil.

(R)-3-(3-Fluorophenoxy)-3-phenylpropylamine (Compound 185)

A solution of (R)-3-(3-fluorophenoxy)-3-phenylpropyl chloride (0.971 g,3.96 mmol), conc. NH₄OH (30 mL), and EtOH (20 mL) were shaken at 90° C.on a Parr apparatus (50-90 psig) for 18 h. The mixture was thenevaporated under vacuum and the residue was dissolved in Et₂O (100 mL)and washed with H₂O (2×25 mL). The organic layer was dried (anh.Na₂SO₄), filtered, and evaporated under vacuum to provide a yellow oil.This material was then dissolved in EtOAc (50 mL) and filtered. Asolution of 30 maleic acid (0.272 g, 2.6 mmol, 0.93 equiv) dissolved inhot EtOAc (5 mL) was added to precipitate the maleate salt (519 mg,53.5%) as a white solid: TLC R_(f) 0.25 (1% MeOH—CHCl₃); GC, t_(r) 7.37min; MS, m/z 245 (M+). The synthesis of Compound 187 was accomplished asfollows.

11-Cyanomethylene-10,11-dihydrodibenzo[b,c]oxepine

To a solution of diethyl cyanomethylphosphonate (5.06 g, 28.6 mmol) indry DMF (15 mL) was added NaH (60% mineral oil dispersion; 1.14 g, 28.5mmol) over a period of 2 min. The reaction was stirred for 10 min andthen a solution of 6,11-dihydrodibenzo[b,c]oxepin-11-one [Kurokawa M. etal., Chem, Pharm, Bull. 39(10), 2564-2573 (1991), hereby incorporated byreference herein] (4.00 g, 19.0 mmol) in dry DMF (5 mL) was added. Thereaction mixture was stirred under argon for 21 h. Water (100 mL) wasthen added and the product was extracted with EtOAc (2×50 mL). Thecombined organic layers were washed with satd. aq. NaCl (2×50 mL), dried(anh. Na₂SO₄), and rotary evaporated. The resulting solid wasrecrystallized from hot EtOAc (10 mL) hexanes (40 mL) to provide 2.43 g(54.7%) of product.

11-Cyanomethyl-11H-10,11-dihydrodibenzo[b,c]oxepine

Following a procedure described in Great Britain Patent 1,129,029 (1968)(Chem, Abstr. 70:37664), hereby incorporated by reference herein], forthe preparation of aluminum amalgam, A1° granules (2.00 g, 74.1 mmol)were first etched with 0.5N NaOH (100 mL) and then washed with H₂O (100mL) followed by EtOH 100 mL). A solution of HgCl₂ (2.00 g, 7.37 mmol) inEt₂O (100 mL) was added. The reaction mixture was stirred for 5 min andthe supernatant was decanted. The solid Al(Hg) amalgam was washed withH₂O (100 mL), EtOH (100 mL), and then Et₂O (100 mL). The amalgam wascovered with Et₂O (100 mL) and a solution of11-cyanomethylene-10,11-dihydrodibenzo[b,c]oxepine (2.00 g, 8.57 mmol)in EtOAc (30 mL) and EtOH (20 mL) was added. Water (2 mL) was added andthe reaction mixture was stirred for 18 h and then filtered. Thefiltrate was rotary evaporated to yield 1.65 g (81.8%) of product as awhite, crystalline solid.

11-(2-Aminoethyl)-11H-10,11-dihydrodibenzo[b,c]oxepine hydrochloride(Compound 187)

To a stirring suspension of lithium aluminum hydride (0.67 g, 18 mmol)in anh. Et₂O (30 mL) was added a solution of11-cyanomethyl-11H-10,11-dihydrodibenzo [b,c]oxepine (1.65 g, 7.01 mmol)in dry THF (5 mL)/anh. Et₂O (10 mL) over a period of 2 min. The reactionwas stirred for 30 min. In the following order, H₂O (0.7 mL), 5N NaOH(0.7 mL), and H₂O (2.1 mL) were added to the reaction mixture. Diethylether (30 mL was added and the mixture was filtered. The filtrate wasrotary evaporated and the resulting oil was dissolved in ETOH (10mL)-Et2O (65 mL). 1.0M HCl in Et₂O (10 mL) was added and the solutionwas allowed to crystallize, giving 1.42 g (73.4%) of the title compound.

Compounds 67-68, 70-75, 79-82, 84-89, 91-95, 98-100, 102, 104-106,109-114, 117, 124-134, 138, and 140-150 were synthesized by standardprocedures known to those skilled in the art, as described above.

Gas Chromatography of Simplified Arylalkylamines

Gas chromatographic and mass spectral data were obtained on aHewlett-Packard-5890 Series II Gas Chromatograph equipped with a 5971Series Mass Selective Detector [Ultra-2 Ultra Performance CapillaryColumn (cross-linked 5% phenyl methyl silicone); column length, 25 m,column i.d., 0.20 mm; The flow rate, 60 mL/min; injector temp., 250° C.;gradient temperature program, 20° C./min from 125 to 325° C. for 10 min,then held constant at 325° C. for 6 min]. Compound 19. (Rt=7.40 min),m/z (rel. int.) 211 (M+,13), 195(16), 194 (100), 193 (73), 180 (8), 179(33), 178 (19) 168 (24), 167 (50), 166 (23), 165 (72), 164 (8), 153(10), 152 (31), 117 (13), 116 (38), 115 (26), 106 (14), 104 (14), 103(24), 102 (8), 91 (11), 78 (14), 77 (29), 63 (9), 51 (17)

Compound 20. (Rt=7.34 min), m/z (rel. int.) 247 (M+,27), 231 (16), 230(100), 229 (45), 215 (29), 214 (14), 204 (43), 203 (37), 202 (13), 201(47), 184 (14), 183 (58), 181 (8), 151 (9), 135 (13), 134 (31), 133(25), 124 (18), 122 (16), 121 (19), 109 (15), 101 (29), 96 (18), 95(11), 83 (11), 75 (20), 57 (10), 42 (9)

Compound 21. (Rt 7.53 min), m/z (rel. int.) 261 (M+,69), 262 (13), 245(17), 244 (100), 230 (11), 229 (42), 216 (11), 215 (15), 214 (14), 204(45), 203 (35), 202 (16), 201 (63), 184. (12), 183 (61), 148 (11), 136(9), 135 (27), 133 (36), 124 (21), 115(16), 109 (43), 83 (12), 74 (8),58 (14), 57 (11)

Compound 22. (Rt=7.37 min), m/z (rel. int.) 261 (M+,4), 244 (14), 229(7), 204 (10), 203 (16), 201 (12), 183 (16), 138 (4), 133 (5), 109 (4),101 (7), 75 (4), 58 (8), 57(4), 44 (100), 42 (7)

Compound 24. (Rt=8.21 min), m/z (rel. int.) 259 (M+,122), 260 (23), 242(44), 241 (15), 228 (15), 227 (49), 216 (15), 213 (56), 212 (16), 211(55), 199 (32), 196 (22), 185 (34), 184 (19), 183 (67), 171 (16), 170(38), 165 (44), 151 (20), 150 (16), 146 (13), 136 (46), 134 (17), 133(37), 123 (15), 121 (22), 120 (13), 109 (100), 91 (34), 77 (29), 51 (15)

Compound 25. (Rt=8.49 min), m/z (rel. int.).259 (M+,39), 243 (16), 242(95), 241 (25), 227 (27), 217 (15), 216 (100), 215 (27), 212 (13), 211(50), 201 (14), 200 (11), 199 (15), 196 (15), 185 (20), 184 (19), 183(50), 171 (24), 170 (28), 165 (15), 146 (10), 136 (11), 134 (12), 133(23), 121 (21), 77 (9).

Compound 26. (Rt=8.69 min), m/z (rel. int.) 259 (M+,11), 243 (17), 242(100), 241 (69), 227 (10), 215 (31), 212 (11), 211 (52), 184 (14), 183(31), 172 (13), 171 (35), 170. (23), 165 (13), 147 (21), 146 (12), 134(19), 133 (23), 121 (13) 91 (11), 77 (10)

Compound 27. (Rt=8.80 min), m/z (rel. int.) 243 (M+,54), 226 (36), 212(12), 211 (69), 200 (14), 199 (16), 198 (20), 197 (100), 196 (39), 185(35), 184 (30), 183 (50), 179 (13), 178 (14), 165 (13), 134 (15), 133(19), 120 (29), 117 (16), 115 (27), 104 (13), 101 (11), 91 (23), 77 (13)

Compound 28. (Rt 8.77 min), m/z (rel. int.) 243 (M+,25), 227 (15), 226(85), 225(26), 212 (19), 211 (100), 200 (22), 199 (17), 197 (18), 196(29), 185 (46), 184 (35), 183 (64), 179 (9), 165 (11), 134 (19), 133(23), 121 (12), 120 (18), 117 (14), 115 (24), 101 (12), 91 (25), 77(12), 65 (11), 51 (9)

Compound 29. (Rt=7.89 min), m/Z (rel. int.) 243 (M+,12), 227 (9), 226(52), 225 (17), 212 (19), 211 (100), 199 (13), 197 (12), 196 (21), 185(19), 184 (24), 183 (43), 179 (7), 134 (11), 133 (15), 120 (9), 117(10), 115 (17), 91 (14)

Compound 30. (Rt=8.36 min), m/z (rel. int.) 263 (M+,21), 246 (26), 220(13), 212 (17), 211 (100), 197 (10), 196 (25), 185 (43), 184 (30), 183(69), 181 (9), 165 (12), 133 (18), 115 (14), 101 (15), 75 (15)

Compound 31. (Rt=9.31 min), m/z (rel. int.) 279 (M+,18), 281 (11), 262(10), 236 (10), 229 (33), 228 (17), 227 (100), 203 (9), 201 (33), 199(15), 192 (15), 178 (19), 166 (18), 165 (53), 164 (13), 163 (16), 140(12), 115 (13), 103 (9)

Compound-32. (Rt=7.30 min), m/z (rel. int.) 229 (M+,21), 213 (16), 212(100), 211 (61), 197 (33), 196 (19), 194 (14), 186 (26), 185 (30), 184(19), 183 (69), 170 (17), 166 (16) 165 (77), 134 (25), 133 (23), 116(17), 115 (17), 103 (18), 101 (11), 78 (13), 77 (23), 75 (13), 51 (18),43 (13), 42 (13)

Compound 33. (Rt 7.56 min), m/z (rel. int.) 261 (M+,68), 245 (18), 244(100), 229 (43), 215 (16), 214 (15), 204 (57), 203 (43), 202 (15), 201(64), 184 (14), 183 (73), 148 (16), 136 (13), 135 (46), 133 (60), 124(51), 115 (27), 111 (14), 109 (96), 107 (16), 96 (14), 83 (27), 75 (20),58 (96), 57 (33), 56 (23), 41 (35)

Compound 34. (Rt=7.39 min), m/z (rel. int.) 261 (M+,72), 262 (14), 245(18), 244 (100), 229 (42), 216 (9), 215 (15), 214 (14), 204 (52), 203(38), 202 (14), 201 (54), 184 (12), 183 (62), 181 (10), 148 (13), 136(9), 135 (31), 133 (40), 124 (30), 115 (18), 109 (57), 107 (9), 83 (13),58 (21), 57 (11)

Compound 35. (Rt=4.45 min), m/z (rel. int.) 181 (M+,8), 165 (10), 164(76), 138 (48) 136 (11), 135 (63), 133 (12), 123 (22), 122 (22), 121(11), 110 (21), 109 (100), 101 (13), 96 (27), 83 (14), 75 (11), 56 (15),45 (21), 44 (40), 42 (9), 41 (15)

Compound 37. (Rt=4.87 min), m/z (rel. int.) 196 (M+,4), 195 (17), 178(76), 163 (20), 152 (41), 150 (22), 137 (12), 136 (29), 135 (60), 133(19), 124 (13), 123 (20), 122 (49), 121 (17), 110 (78), 109 (100), 101(17), 96 (29), 83 (17), 75 (12), 56 (29), 55 (12), 45 (53), 44 (45), 43(39), 41 (30)

Compound 38. (Rt=7.68 min), m/z (rel. int.) 275 (M+,0.1), 203 (5), 201(6), 183 (8), 135 (4), 133 (4), 109 (8), 71 (3), 45 (3), 44 (100), 42(4)

Compound 39. (Rt=7.67 min), m/z (rel. int.) 289 (M+,6), 203 (3), 201(5), 183 (6), 135 (2), 133 (3), 109 (7), 85 (3), 70 (3), 59 (4), 58(100)

Compound 40. (Rt=7.63 min), m/z (rel. int.) 289 (M+,19), 203 (6), 201(13), 183 (17), 152 (5), 135 (6), 133 (8), 109 (15), 85 (5), 70 (4), 58(100)

Compound 41. (Rt=,7.93 min), m/z (rel. int.) 275 (M+,23), 258 (20), 203(27), 202 (14), 201 (52), 184 (9), 183 (59), 181 (10), 150 (11), 149(18), 147 (11), 135 (24), 134 (14), 133 (40), 124(12), 123 (19), 109(76), 107 (9), 103 (10), 83 (15), 75 (10), 72 (100), 71 (12), 57 (18),56 (21), 55 (41)

Compound 43. (Rt=9.18 min), m/z (rel. int.) 293 (M+,11), 276 (10), 243(11), 241 (31), 236 (11), 235 (16), 201 (18), 199 (22), 179 (11), 178(25), 176 (10), 166 (16), 165 (70), 164 (19), 163 (24), 103 (9), 102(9), 75 (11), 44 (100), 43 (11), 42 (15)

Compound 46. (Rt=9.34 min), m/z (rel. int.) 293 (M+,46), 295 (28), 276(16), 243 (24), 242 (15), 241 (75), 237 (12), 236 (18), 201 (33), 199(31), 178 (26), 176 (13), 166 (31), 165 (100), 164 (32), 163 (43), 152(11), 151 (13), 149 (12), 140 (30), 139 (11), 129 (12), 127 (20), 125(31), 117 (26), 116 (26), 115 (64), 91 (12), 89 (17), 77 (13), 75 (22),63 (14), 58 (51), 57 (15), 56 (19), 41 (19)

Compound 50. (Rt=7.37 min), m/z (rel. int.) 261 (M+,2), 244 (9), 229(4), 204 (7), 203 (11), 201 (8), 183 (11), 101 (5), 58 (7), 44 (100), 42(7)

Compound 51. (Rt=7.30 min), m/z (rel. int.) 261 (M+,5), 244 (20), 229(9), 204 (14), 203 (23), 202 (6), 201 (20), 183 (27), 133 (7), 121 (6),101 (9), 75 (6), 58 (7), 44 (100), 43 (6), 42 (11)

Compound 52. (Rt=7.24 min), m/z (rel. int.) 247 (M+,56), 231 (13), 230(81), 229 (47), 216 (12), 215 (32), 214 (16), 204 (29), 203 (31), 202(16), 201 (63), 196 (21), 184 (20), 183 (100), 182 (11), 181 (15), 170(13), 151 (13), 150 (11), 135 (13), 134 (29), 133 (25), 124 (14), 122(20), 121 (21), 109 (13), 101 (27), 96 (21), 75 (23), 43 (14), 42 (15)

Compound 53. (Rt=7.21 min), m/z (rel. int.) 247 (M+,98), 248 (17), 231(13), 230 (84), 229 (56), 215 (38), 214 (16), 203 (33), 202 (16), 201(68), 196 (26), 184 (16), 183 (100), 181 (15), 151 (21), 150(15), 135(14), 134 (35), 133 (24), 124 (19), 122 (23), 121 (25), 111 (13), 101(31), 96 (19), 75 (19)

Compound 55. (Rt 7.86 min) m/z (rel. int.) 275 (M+,98), 276 (20), 258(59), 229 (58), 216 (31), 215 (22), 214 (19), 204 (49), 203 (41), 202(21), 201 (82), 184 (18), 183 (100), 181 (14), 150 (21), 135 (33), 133(55), 124 (41), 115 (13), 109 (90), 101 (15), 83 (20), 75 (16), 72 (23),57 (13), 56 (24)

Compound 56. (Rt=7.79 min), m/z (rel. int.) 261 (M+,67), 262 (12), 244(54), 229 (56), 218 (27), 217 (16), 216 (19), 215 (100), 214 (45), 203(50), 202 (32), 201 (51), 197 (16), 196 (26), 183 (24), 138 (17), 135(20), 134 (17), 133 (39), 122 (26), 121 (13), 109 (30), 101 (17), 96(14), 83 (16), 75(13)

Compound 57. (Rt=7.65 min), m/z (rel. int.) 261 (M+,62) 244 (50), 229(50), 218 (24), 217 (13), 216 (18), 215 (100), 214 (36), 203 (42), 202(19), 201 (33), 197 (14), 196 (19), 183 (17), 138 (19) 135 (16), 134(12), 133 (29), 122 (29), 109 (25), 101 (13)

Compound 58. (Rt=8.15 min), m/z (rel. int.) 275 (M+,134), 276 (26), 258(23), 244 (19), 243 (100), 232. (25), 229 (53), 217 (51), 216 (23), 215(67), 214 (97), 201 (44), 197 (21), 196 (43), 183 (23), 148 (38), 147(21), 138 (46), 135 (46), 134 (18), 133 (64), 125 (25), 123 (28), 122(81), 115 (27), 109 (54), 107 (17), 83 (27), 44 (19), 43 (19)

Compound 59. (Rt=7.61 min), m/z (rel. int.) 275 (M+,27), 204 (8), 203(10), 201 (19), 183 (25), 109 (8), 101 (7), 58 (100), 57 (8), 56 (8), 44(9)

Compound 60. (Rt=7.34 min), m/z (rel. imt.) 261 (M+, 55) 262 (10), 204(16), 203 (15), 201 (31), 183 (35), 133 (11), 122 (11), 121(10), 109(9), 101 (16), 96 (11), 75 (10), 57 (9), 44 (100), 42 (11)

Compound 61. (Rt=8.07 min), m/z (rel. int.) 277 (M+,68) 278 (13), 260(31), 246 (11), 245 (25), 234 (12), 231 (32), 229 (26), 217 (20), 203(23), 201(24), 188 (12), 183 (22), 154 (24), 151 (15), 150 (10), 133(18), 124 (10), 109 (100), 95 (11), 44 (14)

Compound 62. (Rt=8.93 min), m/z (rel. int.) 271 (M+,115), 272 (22), 254(16), 239 (22), 225 (36), 223 (40), 181 (33), 165 (34), 153 (13), 152(24), 136 (39), 132 (13), 131 (16), 123 (20), 122 (13), 121 (89), 119(13), 115 (23), 105 (17), 91 (100), 77 (22)

Compound 63. (Rt=8.47 min), m/z (rel. int.) 287 (M+,31), 241 (9), 204(27), 203 (20), 202 (9), 201 (30), 183 (38), 150 (13), 133 (20), 109(27), 84 (45), 83 (43), 82 (11), 57 (18), 56 (100), 43 (25)

Compound 64. (Rt=8.57 min), m/z (rel. int.) 287 (M+,63), 288 (13), 270(14), 242 (16), 241 (17), 215 (17), 214 (18), 204 (35), 203 (27), 202(18), 201 (70), 183 (86), 150 (18), 147 (16), 146 (17), 135 (16), 133(45), 109 (45), 84 (31), 83 (38), 82 (13), 75 (15), 57 (21), 56 (100),43 (44)

Compound 65. (Rt=8.18 min), m/z (rel. int.) 239 (M+,88), 240 (17), 222(12) 208 (18), 207 (100), 195 (24), 193 (48), 192 (11), 181 (33), 180(32), 179 (57), 178 (72), 166 (16), 165 (60), 152 (13), 130 (36), 129(17), 120 (40), 117 (34), 116 (14), 115 (53), 107 (20), 105 (19), 104(42), 103 (11), 91 (37), 77 (20), 65 (17)

Compound 66. (Rt=7.46 min), m/z (rel. int.) 275 (M+,7), 201 (5), 183(6), 133 (3), 109 (6), 71 (3), 45 (3), 44 (100), 42 (3)

Compound 67. (Rt=7.56 min), m/z (rel. int.) 225 (M+,24), 194 (8), 193(12), 179 (6), 168 (10), 167 (12), 166 (6), 165 (20), 152 (9), 120 (8),116 (6), 115 (7), 103 (7), 77 (8), 51 (5), 44 (100)

Compound 68. (Rt 7.85 min), m/z (rel. int.) 239 (M+,22), 194 (5), 193(10), 168 (10), 167 (12), 166 (6), 165 (19), 152 (9), 134 (6), 116 (5),115 (7), 91 (7), 77 (6), 59 (5), 58 (100), 44 (8)

Compound 69. (Rt=7.35 min), m/z (rel. int.) 275 (M+,11), 203 (24), 202(7), 201 (23), 183 (35), 122 (6), 121 (6), 101 (9), 58 (100), 57 (8), 56(10)

Compound 72. (Rt=7.90 min), m/z (rel. int.) 253 (M+,25), 238 (9), 193(7), 168 (8), 167 (14), 165 (17), 152 (9), 115 (7), 91 (11), 73 (8), 72(100), 58 (45), 56 (7), 44 (6), 43 (9), 42 (8)

Compound 73. (Rt=7.29 min), m/z (rel. int.) 239 (M+,9), 240 (2), 167(2), 165 (5), 152 (2), 115 (2), 77 (2), 59 (5), 58 (100), 44 (3), 42 (5)

Compound 74. (Rt=8.01 min), m/z (rel. int.) 267 (M+,7), 167 (3), 165(6), 152 (3), 91 (4), 87 (7), 86 (100), 72 (13), 58 (10), 56 (4), 42 (4)

Compound 79. (Rt=7.89 min), m/z (rel. int.) 23.0 (M+,37), 214 (15), 213(100), 212 (62), 201 (26), 200 (72), 198 (21), 195 (12), 188 (17), 187(85), 186 (46), 185 (42), 184 (9), 157 (12), 135 (9), 133 (24), 109(10), 107 (20), 106 (62), 80 (14), 79 (32), 78 (9), 51 (20)

Compound 81. (Rt=7.40 min), m/z (rel. int.) 209 (M+,89) 210 (14), 208(100), 193 (17), 192 (56), 191 (42), 189 (12), 178 (20), 166 (11), 165(45), 152 (12), 132 (86), 131 (10), 130 (53), 117 (22), 115 (48), 106(22), 105 (10), 104 (12), 103 (16), 91 (16), 77 (22), 51 (15)

Compound 82. (Rt.=7.93 min), m/z (rel. int.) 275 (M+,124), 276 (25), 232(33), 215 (12), 214 (16), 204 (14), 203 (100), 201 (24), 196 (8), 183(20), 150 (14), 138 (9), 136 (14), 135 (44), 133 (26), 125 (9), 124(71), 123 (29), 121 (14), 115 (14), 111 (72), 110 (9), 109 (84), 101(14), 83 (9), 75 (8)

Compound 83. (Rt=7.22 min), m/z (rel. int.) 235 (M+,10), 219 (17), 218(100), 217 (62), 203 (20), 192 (10), 191 (38), 190 (7), 189 (14), 185(17), 83 (7), 171 (9), 165 (8), 147 (10), 146 (11), 134 (12), 133 (17),121 (8), 109 (8), 97 (8), 45 (7)

Compound 85. (Rt=7.73 min), m/z (rel. int.) 239 (M+,7), 222 (15), 179(8), 178 (9), 168 (16), 167 (33), 166 (12), 165 (43), 161 (9), 152 (20),146 (17), 129 (7), 120 (15), 118 (7), 117 (19), 115 (25), 91 (25), 77(7), 72 (9), 44 (100), 42 (6)

Compound 86. (Rt=7.66 min), m/z (rel. int.) 239 (M+,3), 222 (4), 168(4), 167 (11), 166 (4), 165 (14), 152 (7), 120 (6), 117 (6), 115 (8), 91(9), 72 (5), 44 (100), 42 (3)

Compound 87. (Rt=7.33 min), m/z (rel. int.) 239 (M+,4), 222 (9), 179(9), 178 (11), 168 (11), 167 (27), 166 (13), 165 (48), 161 (7), 152(22), 146 (14), 128 (7), 120 (11), 118 (8), 117 (21), 115 (31), 91 (29),77 (9), 72 (8), 51 (7), 44 (100), 42 (9)

Compound 88. (Rt=7.4 min), m/z (rel. int.) 227 (M+,.O), 183 (10), 168(18), 167 (100), 166 (32), 165 (83), 164 (10), 163 (6), 153 (6), 152(35), 139 (6), 115 (8), 105 (9), 77 (12), 51 (7), 45 (23)

Compound 89. (Rt=8.74 min), m/z (rel. int.) 260 (M+,220), 261 (39), 259(89), 242 (18), 203 (17), 202 (16), 201 (61), 183 (58), 165 (100), 150(20), 148 (25), 138 (24), 137 (61), 122 (73), 121 (31), 111 (47), 101(23), 96 (16), 75 (16), 44 (17), 43 (29)

Compound 90. (Rt=7.32 min), m/z (rel. int.) 235 (M+,9), 219 (16), 218(100), 217 (42), 206 (17), 205 (9), 204 (7), 203 (21), 202 (8), 193(12), 192 (71), 191 (62), 190 (9), 189 (19), 185 (13), 171 (14), 159(9), 147 (14), 146 (16), 134 (10), 133 (17), 121 (14), 109 (11), 101(8), 97 (17), 45 (15)

Compound 91. (Rt=10.67 min), m/z (rel. int.) 329 (M+,6), 301 (20), 300(81), 167 (18), 166 (6), 165 (18), 152 (10), 132 (5), 120 (45), 119(21), 18 (11), 117 (9), 115 (11), 106 (6), 105 (5), 104 (12), 103 (5),92 (8), 91 (100), 77 (10), 41 (6)

Compound 92. (Rt=10.37 min), m/z (rel. int.) 337 (M+,30), 338 (7), 204(7), 203 (7), 201 (7), 183 (10), 133 (6), 121 (8), 120 (70), 106 (6), 92(9), 91 (100)

Compound 93. (Rt=10.25 min), m/z (rel. int.) 351 (M+,28), 352 (7), 337(9), 336 (39), 203 (10), 201 (11), 183 (17), 135 (6), 134 (20), 133 (6),132 (6), 120 (11), 118 (5), 109 (18), 106 (12), 105 (100), 104 (13), 103(8), 91 (14), 79 (11), 77 (12)

Compound 94. (Rt=10.48 min), m/z (rel. int.) 365 (M+,2), 337 (25), 336(100), 203 (8), 201 (8), 183 (14), 133 (5), 132 (6), 120 (14), 119 (13),118 (9), 115 (5), 109 (20), 106 (5), 104 (10), 91 (52)

Compound 95. (Rt=6.68 min), m/z (rel. int.) 283 (M+,59), 284 (11), 267(11), 266 (71), 265 (19), 251 (24), 250 (9), 241 (14), 240 (100), 239(48), 237 (30), 232 (10), 220 (17), 219 (65), 199 (9), 152 (12), 151(18), 142 (20), 140 (13) 139 (20), 127 (22), 119 (24), 114 (12), 101(10), 63 (10), 44 (9)

Compound 96. (Rt=6.93 min), m/z (rel. int.) 265 (M+,46), 249 (16), 248(100), 247 (34), 233 (27), 232 (11), 223 (9) 222 (65), 221 (39), 220(10), 219 (36), 202 (14), 201 (54), 152 (15), 151 (14), 133 (9), 124(12), 119 (9), 109 (9), 101 (14), 75 (9)

Compound 97. (Rt=8.10 min), m/z (rel. int.) 241 (M+,101), 242 (18), 224(50), 223 (19), 210 (11), 209 (37), 197 (12), 196 (10), 195 (55), 194(16), 193 (60), 181 (29), 178 (20), 167 (38), 166 (16), 165 (52), 153(12), 152 (36), 136 (27), 133 (12), 132 (14), 116 (12), 115 (25), 103(13), 91 (100), 77 (18)

Compound 98. (Rt=6.69 min), m/z (rel. int. ) 232 (M+,3), 204 (11), 203(37), 202 (30), 201 (100), 188 (9), 184 (14), 183 (84), 182 (10), 181(15), 170 (9), 109 (17), 107 (10), 83 (10), 75 (8), 57 (7)

Compound 99. (Rt=6. 75 min), m/z (rel. int. ) 233 (M+, 2), 204 (12), 203(68), 202 (26), 201 (100), 200 (6), 188 (9), 184 (13), 183 (86), 182(8), 181 (14), 170 (9), 133 (6), 109 (15), 107 (11), 83 (11), 81 (7), 75(7), 57 (9).

Compound 100. (Rt=7.66 min), m/z (rel. int.) 261 (M+,150), 262 (29), 217(11), 216 (70), 215 (28), 214 (11), 203 (30), 202 (31), 201 (100), 196(10), 184 (15), 183 (90), 181 (11), 133 (20), 124 (12), 122 (20), 109(39), 101 (14), 83 (10), 75 (10), 45 (43)

Compound 101. (Rt=7.72 min), m/z (rel. int.) 245 (M+,20), 229 (16), 228(100), 227 (36), 213 (21), 211 (22), 202 (57), 201 (30), 199 (21), 183(50), 181 (14), 171 (15), 170 (26), 165 (12), 152 (21), 134 (19), 133(35), 122 (28), 120 (19), 120 (13), 119 (12), 109 (20), 107 (20), 106(18), 101 (15), 94 (15), 91 (20), 77 (18), 74 (15), 65 (20), 63 (14), 55(14), 51 (15), 44 (27), 43 (17), 42 (14)

Compound 102. (Rt=8.33 min), m/z (rel. int.) 273 (M+,19), 204 (16), 203(16), 201 (15), 183 (18), 177 (9), 133 (8), 109 (13), 70 (41), 69 (100),68 (20), 43 (25), 42 (5), 41 (5)

Compound 103. (Rt=8.59 min), m/z (rel. int.) 245 (M+,118), 246 (20), 229(15), 228 (100), 227 (85), 213 (27), 211 (23), 209 (15), 207 (12), 202(19), 201 (32), 200 (17), 199 (84), 196 (10), 183 (38), 181 (15), 171(13), 170 (23), 152 (19), 151 (15), 150 (10), 134 (18), 133 (32), 131(12), 122 (36), 119 (15), 109 (24), 107 (10), 106 (12), 91 (19), 77 (12)

Compound 104. (Rt=7.72 min) m/z (rel. int.) 261 (M+,94), 262 (17), 217(15), 216 (92), 215 (18), 204 (12), 203 (86), 202 (25), 201 (100), 184(10), 183 (69), 148 (12), 133 (13), 122 (8), 109 (26), 101. (9) 83 (8),45 (33)

Compound 105. (Rt=10.24 min), m/z (rel. int.) 351 (M+,7), 201 (5), 183(7), 135 (9), 134 (79), 133 (4), 109 (5), 92 (8), 91 (100), 65 (8), 42(7)

Compound 106. (Rt=7.52 min), m/z (rel. int.) 259 (M+,77), 260 (14), 258(31), 244 (30), 228 (13), 227 (28), 214 (14), 201 (24), 165 (12), 164(100), 162 (29), 133 (56), 109 (44), 75 (13), 44 (80), 42 (56)

Compound 107. (Rt=7.45 min), m/z (rel. int.) 227 (M+,101), 228 (16), 226(100), 211 (22), 210 (68), 209 (49), 207 (13), 196 (22), 184 (15), 183(62), 150 (50), 148 (31), 133 (44), 132 (53), 130 (45), 117 (15), 115(29), 106 (14), 77 (18), 75 (13), 51 (14)

Compound 108. (Rt=7.46 min), m/z (rel. int.). 243 (M+,34), 244 (6), 212(6), 211 (9), 197 (6), 186 (12), 185 (10), 184 (5), 183 (19), 165 (15),133 (6), 120 (6), 103 (5), 77 (6), 44 (100), 42 (6)

Compound 109. (Rt=8.68 min), m/z (rel. int.) 285 (M+,110), 286 (22), 284(27), 256 (16), 228 (37), 227 (27), 225 (10), 220 (110), 207 (15), 201(27), 191 (14), 190 (100), 163 (11), 162 (85), 161 (10), 147 (11), 146(11), 133 (32), 109 (20), 83 (12), 82 (36)

Compound 110. (Rt=8.66 min), m/z (rel. int.) 285 (M+,91), 286 (16), 284(100), 243 (16), 227 (26), 225 (11), 221 (10), 220 (17), 214 (12), 207(15), 201 (23), 147 (25), 146 (16), 133 (17), 109 (20), 42 (15)

Compound 111. (Rt=8.81 min), m/z (rel. int.) 287 (M+,29), 214 (9), 204(15), 203 (18), 202 (9), 201 (34), 183 (42), 135 (9), 133 (28), 109(28), 84 (47), 83 (100), 82 (19), 75 (8), 70 (16), 68 (13), 57 (18), 56(28), 44 (16), 43 (25), 42 (14)

Compound 112. (Rt=8.85 min), m/z (rel. int.) 287 (M+,141), 288 (29), 286(22), 202 (21), 201 (62), 183 (64), 133 (23), 109 (27), 84 (100), 83(18), 82 (31), 57 (14), 56 (58), 55 (53), 43 (14), 42 (35)

Compound 113. (Rt=9.08 min), m/z (rel. int.) 251 (M+,27), 180 (38), 179(36), 178 (39), 174 (15), 173 (100), 166 (11), 165 (53), 158 (12), 152(10), 132 (9), 115 (28), 91 (31), 82 (18), 77 (16), 56 (45), 51 (9), 43(23)

Compound 114. (Rt=8.71 min), m/z (rel. int.) 237 (M+,197), 238 (37), 236(67), 193 (15), 179 (30), 178 (40), 165 (41), 159 (43), 158 (26), 132(24), 130 (16), 116 (17), 115 (37), 106 (21), 103 (34), 91 (50), 77(48), 57 (68), 56 (100), 51 (32), 43 (50), 42 (34)

Compound 115. (Rt=9.45 min), m/z (rel. int.) 271 (M+,34), 255 (12), 254(67), 253 (14), 239 (23), 229 (16), 228 (100), 227 (18), 224 (16), 223(68), 213 (9), 212 (10), 211 (10), 197 (34), 196 (17), 195 (11), 181(18), 169 (10), 165 (22), 153 (19), 152 (27), 146 (16), 145 (13), 141(12), 139 (10), 136 (22), 134 (11), 133 (41), 122 (16), 121 (31), 115(30), 91 (18), 77 (15), 65 (11), 63 (10), 44 (10)

Compound 116. (Rt=9.50 min), m/z (rel. int.) 269 (M+,41), 268 (32), 254(8), 253 (21), 252 (100), 251 (14), 238 (23), 237 (18), 221 (10), 209(9), 178 (8), 165 (19), 162 (22), 160 (19), 152 (18), 147 (11), 146 (8),145 (18), 139 (9), 130 (11), 115 (10)

Compound 117. (Rt 7.64 min), m/z (rel. int.) 212 (M+,13), 183 (16), 182(100), 180 (7), 167 (7), 152 (3), 104 (27), 91 (7), 78 (4), 77 (41), 51(13)

Compound 118. (Rt=7.46 min), m/z (rel. int.) 245 (M+,4), 153 (8), 152(43), 150 (9), 135 (6), 133 (10), 124 (5), 123 (36), 122 (38), 121 (17),109 (16), 101 (14), 96 (24), 95 (16), 94 (100), 93 (7), 83 (7), 77 (21),75 (11), 66 (15), 65 (30), 63 (10), 51 (14), 50 (6)

Compound 119. (Rt=7.39 min), m/z (rel. int.) 263 (M+,7), 171 (14), 170(14), 152 (74), 151 (13), 150 (20), 141 (55), 135 (10), 133 (23), 123(20), 122 (100), 121 (49), 120 (11), 113 (9), 112 (92), 111 (9), 109(41), 107 (12), 103 (13),102 (11), 101 (40), 97 (9), 96 (66), 95 (51),94 (9) 84 (28), 83 (88), 82 (8), 81 (16), 77, (14), 75 (54), 74 (10), 70(10), 69 (10), 64 (10), 63 (23), 57 (62), 56 (13), 51 (15), 50 (12), 42(8)

Compound 120. (Rt=8.48 min), m/z (rel. int.) 279 (M+,4), 159 (16), 157(49), 153 (11), 152 (100), 150 (12), 133 (11), 130 (27), 128 (73), 123(12), 122 (57), 121 (23), 111 (10), 109 (25) 101 (23), 99 (16), 96 (26),95 (10), 83 (9), 75 (28), 73 (10), 65 (12), 64 (11), 63 (22), 51 (9), 50(8)

Compound 121. (Rt=8.30 min), m/z (rel. int.) 275 (M+,2), 152 (15), 125(8), 124 (100), 122 (14), 121 (7), 109 (35), 96 (7), 95 (10), 81 (14),77 (9), 65 (7), 52 (11)

Compound 122. (Rt=7.39 min), m/z (rel. int.) 263 (M+,0.1), 170 (12), 152(66), 151 (10), 150 (18), 141 (68), 135 (10), 133 (19), 123 (16), 122(76), 121 (39), 112 (100), 111 (18), 109 (36), 107 (11), 103 (11), 102(9), 101 (33), 96 (56), 95 (32), 92(11), 83 (96), 81 (13), 77 (13), 75(43), 64 (25), 63 (26), 57 (61), 56 (14), 51 (14), 50 (11)

Compound 123. (Rt=5.88 min), m/z (rel. int.) 275 (M+,46), 276 (9), 202(8), 201 (30), 183 (28), 133 (8), 109 (9), 101 (9), 71 (9), 59 (12), 58(100), 44 (8), 42 (26)

Compound 124. (Rt=7.05 min), m/z (rel. int.) 229 (M+,15), 213 (15), 212(89), 211 (13), 198 (20), 197 (100), 196 (24), 186 (12), 185 (21), 184(29), 183 (87), 179 (7), 178 (8), 177 (13), 176 (5), 171 (7), 170 (18),169 (4), 166 (5), 165 (20), 152 (5), 133 (7), 75 (4), 63 (4), 57 (9), 56(4)

Compound 125. (Rt=7.54 min), m/z (rel. int.) 225 (M+,57), 226 (13), 209(13), 208 (75), 193 (13), 180 (14), 179 (21), 178 (20), 165 (22), 130(34), 117 (59), 115 (29), 105 (18), 104 (94), 103 (45), 91 (100), 78(30), 77 (38), 65 (36), 63 (13), 51 (20), 45 (17)

Compound 126. (Rt 7.81 min), m/z (rel. int.) 261 (M+,12), 244 (31), 152(27), 151 (17), 150 (9), 136 (11), 135 (100), 133 (21), 122 (24), 115(9), 110 (13), 109 (90), 107 (6), 96 (7), 83 (27), 56 (7)

Compound 127. (Rt=7.93 m/z), m/z (rel. int.) 225 (M+,23), 208 (20), 207(6), 193 (13), 181 (7), 180 (37), 179 (100), 178 (36), 167 (9), 166(12), 165 (36), 152 (9), 134 (30), 130 (26), 129 (9), 117 (18), 115(22), 104 (6), 91 (38), 77 (7), 65 (7)

Compound 128. (Rt 7.42 min), m/z (rel. int.) 211 (M+,83), 212 (15), 194(36), 193 (18), 182 (62), 181 (20), 180 (17), 179 (53) 178 (60), 176(11), 167 (57), 166 (44), 165 (100), 152 (24), 120 (39), 116 (12), 115(28), 104 (22), 103 (15), 91 (46), 89 (16), 78 (10), 77 (20), 65 (15),63 (12), 51 (12)

Compound 129. (Rt=7.39 min) m/z (rel. int.) 229 (M+,104), 230 (19), 212(28), 211 (14), 201 (13), 200 (85), 199 (22), 198 (14), 197 (50), 196(58), 185 (73), 184 (45), 183 (100), 179 (43), 178 (55), 177 (17), 176(17), 170 (18), 165 (33), 152 (12), 133 (22), 120 (57), 115 (17), 109(44), 104 (23), 103 (17), 91 (32), 89 (16), 83 (20), 78 (12), 77 (22),63 (16), 51 (13)

Compound 130. (Rt=7.38 min), m/z (rel. int.) 229 (M+,133) 230 (24), 212(27), 211 (14), 200 (54), 199 (17), 198 (16), 197 (53), 196 (64), 185(49), 184 (43), 183 (100), 179 (28), 178 (29), 177 (14), 170 (19), 165(26), 133 (22), 120 (35), 115 (19), 109 (32), 104 (17), 103 (18), 91(38), 89 (17), 83 (18), 77 (24), 63 (16)

Compound 131. (Rt=7.40 min), m/z (rel. int.) 229 (M+,146), 230 (26), 212(48), 211 (23), 200 (51), 199 (17), 198 (16), 197 (61), 196 (70), 185(50), 184 (43), 183 (100), 179 (28), 178 (28), 170 (20), 165 (23), 133(21), 120 (35), 115 (20), 109 (59), 104 (25), 103 (17), 91 (27), 89(17), 83 (22), 77 (22)

Compound 132. (Rt=7.03 min), m/z (rel. int.) 0 (M+,.O), 185 (14), 184(100), 183 (23), 181 (17), 165 (18), 155 (12), 153 (14), 152 (12), 120(85), 119 (67), 115 (10), 106 (16), 91 (19), 89 (14), 78 (12), 77 (25),51 (16)

Compound 133. (Rt=7.09 min), m/z (rel. int.) 211 (M+,13), 195 (16), 194(100), 181 (27), 180 (70), 179 (31), 178 (28) 166 (25), 165 (40), 152(9), 120 (14), 119 (14), 118 (102), 115 (10), 104 (26), 103 (53), 102(12), 91 (62), 89 (′-O), 78 (13), 77 (42), 65 (17), 51 (13)

Compound 134. (Rt=7.45 min), m/z (rel. int.) 211 (M+,14), 183 (15), 182(100), 181 (14), 179 (13), 178 (18), 167 (27) 166 (18), 165 (46), 152(10), 115 (8), 104 (8), 103 (6), 91 (29), 89 (7), 78 (5), 77 (7), 65 (7)

Compound 135. (Rt=8.60 min), m/z (rel. int.) 273 (M+,34), 257 (14), 256(76), 231 (16), 230 (100), 228 (18), 227 (57), 213 (14), 211 (37), 202(30), 201 (40), 199 (26), 184 (13), 183 (50), 151 (12), 171 (17), 170(20), 152 (15), 150 (19), 134 (15), 133 (31), 122 (14), 121 (29), 109(16), 107 (13), 106 (17), 91 (12), 65 (12)

Compound 136. (Rt 9.26 min), m/z (rel. int.) 275 (M+,44), 277 (15), 260(28), 259 (19), 258 (81), 257 (13), 243 (15), 234 (33), 233 (19), 232(100), 231 (13), 229 (15), 227 (42), 224 (15), 223 (86), 208 (13), 197(45), 196 (26), 195 (13), 182 (14), 181 (33), 179 (11) 178 (18), 166(22), 165 (60), 164 (12), 163 (10), 153 (32), 152 (55), 151 (18), 149(10), 139 (11), 137 (17), 136 (19), 121 (13), 115 (25), 102 (11), 91(16), 77 (17)

Compound 137. (Rt=7.42 min), m/z (rel. int.) 245 (M+,1), 153. (8) 152(7), 141 (64), 135 (10), 134 (100), 132 (11), 117 (6), 115 (12), 112(56), 105 (15), 104 (55), 103 (32), 95 (8), 91 (16), 84 (8), 83 (15), 78(24), 77 (24), 75 (9), 65 (6), 63 (8), 57 (10), 51 (9)

Compound 138. (Rt=9.24 min), m/z (rel. int.) 289 (M+,77), 290 (16), 230(20), 229 (21), 215 (15), 203 (22), 201 (32), 183 (36), 134 (10), 133(13), 124 (10), 121 (9), 109 (10), 101 (10), 73 (100), 43 (23)

Compound 139. (Rt=7.25 min), m/z (rel. int.) 245 (M+,92), 246 (15), 244(67), 229 (16), 228 (63), 227 (46), 225 (10), 224 (15), 214 (13), 201(39, 183 (13), 151 (13), 150 (100), 149 (14), 148 (58), 135 (22), 133(54), 124 (14), 122 (12), 109 (18), 101 (15), 75 (13)

Compound 140a. (Rt=8.64 min), m/z (rel. int.) 271 (M+,72), 272 (14), 270(37), 255 (21), 254 (100), 242 (19), 227 (14), 226 (63), 225 (50), 199(19), 197 (30), 196 (25), 183 (32), 176 (27), 170 (20), 150 (44), 148(34), 146 (14), 133 (32), 131 (14), 121 (11)

Compound 140b. (Rt=8.68 min), m/z (rel. int.) 271 (M+,57), 272 (10), 270(33), 255 (20), 254 (100), 242 (15), 227 (12), 226 (54), 225 (40), 209(8), 199 (14), 197 (22), 196 (19), 183 (25), 176 (21), 170 (16), 150(33), 148 (22), 146 (9), 133 (20), 131 (10)

Compound 141. (R_(t)=8.44 min), m/z (rel. int.) 257 (M+,48), 258 (8),256 (36), 241 (21), 240 (100), 239 (19), 226 (22), 225 (20), 209 (11),197 (14), 196 (18), 183 (25), 170 (16), 162 (19), 160 (10), 150 (28),148 (26), 147 (9), 146 (8), 145 (13), 133 (20), 130 (8), 121 (10)

Compound 142. (R_(t)=8.47 min), m/z (rel. int.) 273 (M+,14), 217 (5),216 (31), 215 (5), 183 (8), 170 (4), 150 (5), 121 (4), 58 (5), 45 (5),44 (100)

Compound 143. (R_(t)=9.39 mini m/z (rel. int.) 273 (M+,47), 275 (16),274 (19) 272 (36), 258 (39), 257 (26), 256 (100), 255 (17), 242 (25),241 (15), 221 (23), 178 (25), 177 (11), 176 (14), 168 (14), 167 (11),166 (54), 165 (34), 164 (34), 163 (16), 162 (45), 160 (19), 152 (28),151 (22), 149 (19) 147 (18), 145 (24), 139 (11), 136 (15), 131 (15),130(35), 121 (15), 115 (14), 111 (11), 103 (13), 102 (19) 89 (11), 77(16), 75 (14), 63 (16), 51 (12)

Compound 145. (Rt=7.35 min), m/z (rel. int.) 277 (M+,7), 122 (10), 109(8), 96 (6), 95 (6), 83 (10), 75 (6), 63 (2), 57 (7), 44 (100), 42 (9).

Compound 148. (Rt=8.43 min), m/z (rel. int.) 261 (M+,3), 170 (14), 169(5), 168 (44), 153 (4), 151 (4), 140 (6), 139 (4), 138 (15), 132 (6),125 (7), 123 (40), 115 (6), 103 (24), 102 (8), 101 (5), 95 (7), 94(100), 89 (5), 77 (22), 75 (6), 66 (S), 65 (16), 63 (7), 51 (10), 50 (4)

Compound 149. (Rt,=9.28 min), m/z (rel. int.) 295 (M+,4), 170 (32), 169(12), 168 (100), 166 (8), 159 (22), 157 (66), 152 (11), 140 (16), 139(11), 138 (41), 132 (11), 130 (32), 129 (8), 128 (82), 127 (10), 125(16), 115 (12), 111 (15), 103 (55), 102 (18) 101 (15), 99 (19), 89 (10),77 (26), 76 (8), 75 (27), 73 (11), 65 (11), 64 (10), 63 (22), 51 (11)

Compound 150. (Rt=8.32 min), m/z (rel int.) 279 (M+,4), 171 (9), 170(37), 169 (13), 168 (100), 166 (8), 142 (8), 141 (88), 140 (19), 139(12), 138 (42), 132 (12), 130 (7), 125 (16), 115 (12), 113 (10), 112(89), 111 (11), 104 (8), 103 (60), 102 (19), 101 (12), 95 (14), 89 (11),84 (11), 83 (24), 77 (29), 76 (6), 75 (24), 63 (13), 57 (17), 51 (11).

Compound 151. (Rt=7.68 min), m/z (rel. int.) 261 (M+,62), 244 (8), 216(79), 203 (65), 201 (82), 183 (100), 121 (50), 101 (40), 75 (35), 44(52).

Compound 152. (Rt=8.097 min), m/z (rel. int.) 42 (34), 43 (10), 44 (42),56 (13), 57 (10), 58 (72), 71 (6), 72 (74), 73 (14), 74 (14), 75 (14),86 (15), 95 (9), 96 (10), 100 (42), 101 (31), 114 (90), 115 (8), 120(10), 121 (20), 122 (9), 123 (7), 138 (10), 149 (9), 164 (7), 170 (8),181 (7), 183 (100), 184 (15), 188 (8), 194 (6), 195 (7), 196 (10), 201(64), 202 (21), 203 (63), 204 (10), 214 (12), 215 (12), 216 (12), 317(92), 318 (20).

Compound 153. (Rt=7.88 min), m/z (rel. int.) 42 (7), 43 (8), 44 (32), 46(16), 72 (24), 75 (11), 86 (21), 95 (8), 96 (9), 101 (20), 109 (14), 121(30), 122 (14), 123 (6), 139 (6), 149 (10), 170 (6), 181 (6), 183 (59),184 (9), 188 (6), 194 (11), 195 (7), 196 (10), 201 (51), 202 (17), 203(56), 204 (8), 214 (10), 215 (18), 216 (52), 217 (13), 289 (100), 290(20)

Compound 154. (Rt=7.74 min), m/z (rel. int.) 42 (16), 44 (23), 45 (35),46 (15), 58 (20), 72 (59), 73 (17), 75 (12), 86 (23), 96 (9), 101 (18),121 (22), 183 (52), 194 (9), 201 (44), 202 (15), 203 (41), 214 (9), 215(11), 216 (20), 217 (10), 289 (100), 290 (20).

Compound 155. (Rt=7.67 min), m/z (rel. int.) 58 (44), 75 (15), 95 (9),96 (11), 101 (22), 109 (16), 121 (33), 122 (15), 125 (12), 149 (9), 183(62), 184 (10), 196 (12), 201 (57), 202 (19), 203 (53), 214 (11), 215(19), 217 (13), 275 (100), 276 (18)

Compound 156. (Rt=8.93 min), m/z (rel. int.) 237 (M+,11), 220 (41), 219(30), 219 (30), 206 (7), 205 (39), 204 (5), 194 (28), 193 (100) 192(21), 191 (31), 190 (9), 189 (17), 179 (15), 178 (50), 177 (5), 176 (5),165 (20), 152 (7), 128 (5), 116 (7), 115 (39), 91 (11)

Compound 157. (Rt=10.88 min), m/z (rel. int.) 343 (M+,6), 300 (100), 167(16), 166 (6), 166 (6), 165 (16), 152 (9), 133 (8), 120 (28), 118 (8),117 (6), 115 (8), 104 (11), 92 (5), 91 (62), 77 (7).

Compound 158. (Rt=10.74 min), m/z (rel. int.) 342 (M+,0.0) 300 (100),167 (13), 166 (5), 165 (13), 152 (7), 120 (22), 118 (6), 115 (6), 106(3), 104 (7), 91 (31)

Compound 159. (Rt=11.41 min), m/z (rel. int.) 363 (M+,13), 336 (33), 335(24), 334 (95), 182 (10), 181 (9), 168 (14), 167 (40), 166 (18), 165(36), 156 (27), 155 (15), 154 (85), 153 (27), 152 (29), 140 (9), 139(7), 138 (14), 127 (32), 126 (9), 125 (100), 117 (19), 116 (7), 115(24), 103 (11), 91 (43), 77 (15), 72 (7), 41 (12).

Compound 160. (Rt=11.48 min.), m/z (rel. int.) 363 (M+,8) 336 (35), 335(25), 334 (100), 182 (5), 181 (11), 168 (9), 167 (29), 166 (13), 165(29), 156 (11), 155 (11), 154 (37), 153 (26), 152 (23), 140 (8), 139(6), 138 (12), 127 (26), 126 (7), 125 (81), 125 (81), 117 (13), 115(17), 103 (7), 91 (26), 89 (5), 77 (9).

Compound 161. (Rt=11.83 min), m/z (rel. int.) 408 (M+,4), 407 (12), 381(24), 380 (98), 379 (25), 378 (100), 200 (77), 199 (31), 198 (87), 197(24), 184 (17), 182 (15), 181 (16), 171 (75), 169 (77), 168 (18), 167(60), 166 (22), 165 (58), 152 (32), 118 (27), 117 (47), 116 (13), 115(37), 104 (13), 103 (19), 91 (64), 90 (17), 77 (25)

Compound 162. (Rt=12.02 min), m/z (rel. int.) 408 (M+,3), 380 (100), 379(25), 378 (99), 200 (40), 199 (32), 198 (48), 197 (30), 184 (18), 182(16), 181 (23), 171 (83), 169 (85), 168 (16), 167 (49), 166 (18), 165(50), 152 (28), 119 (11), 118 (32), 117 (46), 116 (12), 115 (34), 104(11), 103 (17), 91 (63), 90 (16), 89 (10), 77 (23).

Compound 163. (Rt=10.58 min,), m/z (rel. int.) 347 (M+,14), 318 (100),181 (5), 168 (6), 167 (24), 166 (17), 165 (26), 152 (16), 150 (6), 139(8), 138 (66), 137 (23), 137 (23), 136 (10), 124 (6), 122 (13), 117 (8),115 (14), 110 (8), 109 (100), 103 (6), 91 (22), 77 (7).

Compound 164. (Rt=10.59 min), m/z (rel. int. 347 (M+,14) 318 (100), 181(5), 178 (5), 168 (7), 167 (27), 166 (17), 165 (28), 152 (16), 150 (6),139 (8), 138 (70), 137 (18), 136 (11), 122 (14), 117 (6), 115 (13), 110(7), 109 (79), 103 (6), 91 (20), 91 (20), 77 (6).

Compound 165. (Rt=10.61 min), m/z (rel. int. 347 (M+,8), 318 (95), 181(8), 167 (20), 166 (10), 165 (21), 152 (13), 138 (34), 137 (27), 136(11), 136 (11), 122 (14), 117 (6), 115 (11), 110 (8), 109 (100), 91(22), 77 (6).

Compound 166. (Rt=11.62 min), m/z (rel. int.) 359 (M+,2), 330 (100), 167(13), 165 (14), 152 (8), 150 (6), 149 (38), 148 (7), 135 (8), 134 (13),122 (7), 122 (7), 121 (73), 117 (6), 115 (8), 91 (23), 77 (7)

Compound 167. (Rt=11.18 min) m/z (rel. int.) 359 (M+,4) 330 (100), 136(0) 121 (6).

Compound 168. (Rt=10.86 min) m/z (rel. int.) 343 (M+,6) 314 (100), 167(16), 166 (6), 165 (16), 152 (9), 134 (17.), 133 (13), 132 (6), 118(14), 117 (9), 115 (10), 106 (7), 106 (7), 105 (59), 91 (20), 77 (6).

Compound 169. (Rt=10.94 min), m/z (rel int. 343 (M+,4), 314 (100), 167(14), 166 (5), 165 (15), 152 (8), 134 (9), 133 (16), 132 (7), 732 (7),118 (14), 117 (8), 115 (9), 106 (6), 105 (62), 91 (18), 77 (5).

Compound 170. (Rt=12.52 min), m/z (rel. int.) 374 (M+,13), 345 (50), 315(5), 207 (12), 194 (8), 193 (16), 179 (5), 168 (9), 167 (22), 166 (16),165 (100), 164 (15), 164 (15), 152 (12), 136 (45), 117 (13), 115 (11),104 (6), 103 (6), 91 (26), 90 (8), 77 (7).

Compound 171. (Rt=11.16 min), m/z (rel. int.) 341 (M+,14), 182 (8), 181(53), 168 (6), 167 (18), 167 (18), 166 (8), 165 (21), 152 (11), 144 (8),132 (18), 131 (100), 129 (10), 128 (5), 120 (12), 118 (5), 117 (9), 116(10), 115 (15), 106 (5), 104 (9), 103 (9), 91 (48), 77 (10).

Compound 172. (Rt=8.53 min), m/z (rel. int.) 275 (M+,165), 274 (95), 260(18), 259 (24), 258 (87), 257 (28), 254 (17), 244 (41), 243 (46), 242(18), 233 (30), 214 (26), 201 (46), 189 (18), 188 (36), 183 (27), 181(22), 180 (100), 178 (24), 165 (38), 163 (28), 154 (17), 150 (39), 149(25), 148 (82), 139 (58), 135 (20), 133 (35), 109 (20).

Compound 173. (Rt=8.44 min), m/z (rel. int.) 277 (M+,11), 260 (80), 259(34), 245 (8), 241 (7), 234 (8), 233 (23), 230 (19), 229 (100), 214 (9),203 (12), 202 (17), 201 (25), 190 (8), 189 (25), 188 (18), 183 (18), 171(7), 170 (15), 165 (14), 164 (7), 154 (7), 152 (7), 151 (8), 134 (12),133 (23), 121 (7), 109 (13), 101 (8).

Compound 174. (Rt=8.49 min), m/z (rel. int.) 291 (M+,59), 260 (27), 259(17), 234 (18), 233 (14), 230 (10), 229 (55), 203 (12), 202 (10), 201(18), 189 (20), 188 (14), 183 (15), 170 (12), 169 (6), 168 (13), 165(14), 164 (8), 152 (8), 151 (6), 138 (7), 137 (8), 134 (7), 133 (13),109 (12), 101 (7) 57 (6), 44 (100), 42 (6).

Compound 175. (Rt=9.64 min), m/z (rel. int.) 303 (M+,8), 123 (47), 109(6), 96 (7), 95 (40), 85 (26), 84 (100), 82 (7), 75 (9), 68 (4), 56(25), 55 (29), 43 (9), 42 (16).

Compound 176. (Rt=8.35 min), m/z (rel. int.) 277 (M+,24), 245 (8), 220(6), 219 (5), 183 (4), 171 (6), 170 (10), 151 (5), 138 (7), 109 (5), 57(6), 44 (100), 42 (6).

Compound 177. (Rt=7.83 min), m/z (rel. int.) 241 (M+,0.1), 134 (30), 109(8), 108 (100), 107 (16), 104 (17), 103 (9), 91 (12), 90 (4), 79 (10),78 (14), 77 (24), 65 (8) 51 (9).

Compound 178. (Rt=8.29 min), m/z (rel. int.) 257 (M+,0.1), 134 (17), 125(8), 124 (100), 109 (23), 104 (10), 103 (6), 95 (4), 91 (5), 81 (9), 78(7), 77 (11), 65 (7), 52 (9), 51 (6)

Compound 179. (RC 7.88 min), m/z (rel. int.) 255 (M+,6), 148 (12), 115(5), 108 (8), 107 (7), 104 (12), 103 (7), 91 (11), 79 (5), 78 (11), 77(19), 65 (6), 51 (5), 44 (100), 42 (8).

Compound 180. (Rt=7.28 min), m/z (rel. int.) 295 (M+,1), 183 (10), 162(22), 145 (11), 143 (28), 135 (10), 134 (100), 133 (11), 132 (13), 117(6), 115 (12), 114 (6), 113 (9), 112 (7), 105 (17), 104 (57), 103 (32),102 (6), 95 (7), 91 (18), 89 (5), 83 (8), 79 (6), 78 (28), 77 (28), 75(6), 65 (8), 63 (11), 51 (11)

Compound 181. (Rt=7.7 min), m/z (rel. int.) 259 (M+,3), 137 (16), 135(5), 122 (15), 121 (6), 109 (15), 108 (100), 107 (18), 96 (7), 91 (5),79 (7), 78 (5), 77 (17), 65 (5), 51 (5).

Compound 182. (Rt=8.00 min), m/z (rel. int.) 225 (M+,2), 208 (51), 207(40), 182 (14), 181 (100), 165 (7), 152 (24), 151 (8), 74 (2).

Compound 183. (Rt=8.98 min), m/z (rel. int.) 241 (M+,7), 224 (33), 223(46), 199 (6), 198 (15), 197 (100), 178 (6), 165 (28), 152 (13) 150 (2).

Compound 184. (Rt=8.90 min), m/z (rel. int.) 235 (M+,3), 218 (30), 217(13), 203 (8), 202 (9), 193 (10), 192 (67), 191 (100), 190 (11), 189(29), 178 (7), 168 (18), 152 (3)

Compound 185. (Rt=7.37 min), m/z (rel. int.) 245 (M+,0.1), 152 (8), 141(61), 135 (10), 134 (100), 132 (11), 115 (14), 112 (64), 105 (19), 104(71), 103 (45), 102 (8), 95 (13), 91 (24), 89 (8), 84 (12), 83 (28), 79(10), 78 (42), 77 (44), 75 (16), 65 (12), 64 (7), 63 (16), 57 (24), 56(7), 52 (7) 51 (22), 50 (9).

Compound 186. (Rt=7.31 min), m/z (rel. int.) 245 (M+,0.1), 152 (8), 141(64), 135 (10), 134 (100), 132 (11), 117 (6), 115 (13), 112 (38), 77(40), 75 (14), 65 (11), 64 (6), 63 (14), 57 (21), 52 (6), 51 (18), 50(7).

Compound 187. (Rt=8.64 min), m/z (rel. int.) 239 (M+,0.0), 221 (17), 220(6), 207 (8), 196 (11), 195 (39), 194 (14), 193 (11), 192 (38), 191 (7),181 (6), 179 (21), 178 (100), 168 (7), 167 (41), 166 (17), 165 (53), 164(6), 153 (6), 152 (26), 139 (6), 128 (7), 115 (18), 91 (6), 89 (6), 77(6), 63 (6), 51 (5), 44 (6).

EXAMPLE 30 Biological Properties of Synthesized Arylalkylamines

Compounds synthesized as described in Example 28 and Example 29 weretested for various biological properties detailed in the examples. TABLE1 IC₅₀ (μM) IC₅₀ μM) vs. Compound vs. NMDA^(a) [3H]MK-801c Compound 10.102 (7) 126 (4) Compound 2 0.192 (4) not tested Compound 3 0.003 (7)not tested Compound 4 0.184 (5) 89 (1) Compound 5 0.102 (1) 15.2 (2)0.070 (3)^(b) Compound 6 0.129 (1) >100 (1) (0% at 100 μm)^(d) Compound7 0.163 (2) 129 (1) Compound 8 0.099 (2) 219 (1) Compound 9 1.2 (5) >100(2) (10% at 100 μm)^(d) Compound 10 0.082 (2) ˜80 (1) (57% at 80 μm)^(d)Compound 11 4.0 (2) not tested Compound 12 6.0 (11) 98 (1) Compound 13not tested not tested Compound 14 8.8 (2) ˜100 μm Compound 15 4.9 (3)˜100 μm Compound 16 5.1 (1) 28.8 (1) Compound 17 9.6 (1) 36.3 (1)Compound 18 5.1 (3) 34 (1) Compound 19 0.435 (11) 2.1 (5) Compound 200.070 (15) 0.252 (9) Compound 21 0.038 (3) 0.457 (2) Compound 22 0.145(6) 3.45 (2) Compound 23 0.267 (3) 5.4 (1) Compound 24 0.206 (6) 0.591(6) Compound 25 0.279 (2) 0.871 (2) Compound 26 27 (2) 34 (2) Compound27 0.071 (1) 0.180 (2) Compound 28 0.380 (1) 2.3 (3) Compound 29 1.9 (2)5.8 (3) Compound 30 0.035 (2) 0.407 (2) Compound 31 0.052 (7) 1.3 (2)Compound 32 0.284 (5) 0.799 (3) Compound 33 0.060 (9) 0.181 (6) Compound34 0.426 (6) 2.7 (3) Compound 35 6.2 (1) 25.1 (1) Compound 36 not testednot tested Compound 37 0.944 (2) 11.1 (2) Compound 38 0.407 (2) 2.3 (2)Compound 39 0.251 (1) 2.9 (3) Compound 40 0.933 (1) 18.1 (3) Compound 410.724 (1) 14.0 (3) Compound 42 not tested not tested Compound 43 0.232(4) 7.5 (2) Compound 44 not tested not tested Compound 45 not tested nottested Compound 46 0.013 (3) 5.2 (2) Compound 47 not tested not testedCompound 48 not tested not tested Compound 49 not tested not testedCompound 50 0.089 (6) 0.762 (4) Compound 51 1.1 (4) 4.5 (2) Compound 520.102 (3) 0.380 (2) Compound 53 0.217 (3) 4.2 (2) Compound 54 0.036 (4)0.046 (3) Compound 55 0.035 (3) 0.153 (2) Compound 56 0.218 (4) 0.955(2) Compound 57 0.028 (4) 0.063 (2) Compound 58 0.028 (2) 0.203 (3)Compound 59 0.272 (2) 0.453 (3) Compound 60 0.416 (11) 0.641 (9)Compound 61 0.134 (4) 0.324 (2) Compound 62 0.177 (5) 0.617 (1) Compound63 0.093 (6) 0.245 (3) Compound 64 0.309 (3) 0.851 (2) Compound 65 0.167(3) 2.0 (1) Compound 66 0.236 (4) 1.2 (2) Compound 67 10.95 (2) 2.9 (1)Compound 68 2.9 (1) not tested Compound 69 0.224 (2) 0.366 (1) Compound70 1.7 (1) not tested Compound 71 6.35 (2) not tested Compound 72 7.4(1) not tested Compound 73 12.6 (1) not tested Compound 74 27.5 (1) nottested Compound 75 0.94 (2) not tested Compound 76 0.73 (2) not testedCompound 77 5.5 (2) not tested Compound 78 10.2 (1) not tested Compound79 12.6 (4) 10.2 (2) Compound 80 28 (1) 182 (1) Compound 81 1.4 (1) 6.1(2) Compound 82 0.106 (5) 0.794 (1) Compound 83 0.341 (4) 0.794 (1)Compound 84 7.9 (2) 23.4 (1) Compound 85 1.2 (3) 3.5 (1) Compound 86 1.2(3) 6.0 (1) Compbund 87 0.657 (4) 3.0 (1) Compound 88 2.5 (3) 10.6 (2)Compound 89 0.240 (3) 1.2 (2) Compound 90 0.270 (4) 1.4 (2) Compound 910.162 (3) 14.1 (2) Compound 92 1.3 (3) 20.2 (2) Compound 93 0.486 (3)26.9 (2) Compound 94 0.248 (4) 22.6 (2) Compound 95 0.311 (3) 3.0 (2)Compound 96 0.187 (5) 1.1 (2) Compound 97 0.410 (3) 2.6 (1) Compound 987.9 (1) 52.5 (2) Compound 99 >100 (1) 105 (2) Compound 100 0.602 (2) 3.2(1) Compound 101 0.912 (2) 2.0 (1) Compound 102 1.01 (2) 3.3 (1)Compound 103 0.380 (4) 0.661 (2) Compound 104 7.983 (3) >10 (1) Compound105 1.03 (1) >3 (1) Compound 106 0.767 (1) 1.31 (1) Compound 107 2.67(1) 3.83 (1) Compound 108 1.06 (1) 0.942 (1) Compound 109 1.95 (1) 1.08(3) Compound 110 42.7 (1) 13.3 (1) Compound 111 0.645 (3) 0.167 (2)Compound 112 28.0 (2) 21.0 (1) Compound 113 13.5 (1) not tested Compound114 3.4 (1) not tested Compound 115 1.4 (1) 1.0 (1) Compound 116 3.6 (1)not tested Compound 117 19.6 (2) 6.0 (2) Compound 118 0.409 (2) 0.240(3) Compound 119 0.115 (4) 0.087 (3) Compound 120 0.101 (3) 0.074 (3)Compound 121 0.656 (3) 0.670 (3) Compound 122 0.209 (2) 0.342 (2)Compound 123 9.6 (7) >3 (2) Compound 124 3.5 (1) 14.3 (3) Compound 1251.7 (1) 6.7 (2) Compound 126 0.398 (3) 6.0 (1) Compound 127 1.2 (3) 17.5(2) Compound 128 0.646 (4) 5.5 (1) Compound 129 1.26 (2) not testedCompound 130 0.851 (2) not tested Compound 131 1.23 (2) not testedCompound 132 1.3 (1) 6.4 (1) Compound 133 0.760 (1) 3.0 (1) Compound 1342.5 (1) >10 (1) Compound 135 0.244 (2) 1.185 (2) Compound 136 0.139 (2)0.706 (1) Compound 137 0.232 (3) 0.074 (2) Compound 138 107 (1) >100 (1)Compound 139 1.97 (2) 5.6 (2) Compound 140 20.8 (1) not tested Compound141 4.26 (1) 8.97 (1) Compound 142 1.013 (3) 1.54 (2) Compound 143 2.82(1) not tested Compound 144 not tested not tested Compound 145 0.098 (1)0.626 (1) Compound 146 0.829 (3) 0.372 (1) Compound 147 0.894 (2) nottested Compound 148 0.549 (2) 0.373 (2) Compound 149 0.085 (3) 0.150 (3)Compound 150 0.195 (2) 0.351 (2) Compound 151 54.9 (1) >100 (1) Compound152 not tested not tested Compound 153 not tested not tested Compound154 not tested not tested Compound 155 not tested not tested Compound156 0.069 (3) 0.090 (2) Compound 157 0.142 (2) 23.16 (2) Compound 1580.351 (2) 39.64 (1) Compound 159 0.185 (2) 10.41 (1) Compound 160 7.35(3) 48.94 (1) Compound 161 0.247 (2) 5.62 (1) Compound 162 1.138 (2)76.41 (1) Compound 163 0.326 (2) 10.34 (1) Compound 164 0.475 (2) 18.30(1) Compound 165 0.337 (2) 171 (1) Compound 166 0.619 (2) 36.7 (1)Compound 167 0.080 (2) 14.5 (1) Compound 168 0.092 (2) 17.4 (1) Compound169 0.298 (2) 26.7 (1) Compound 170 0.238 (2) 57.0 (1) Compound 1710.310 (3) 39.6 (1) Compound 172 38.0 (1) 37.3 (1) Compound 173 22.9 (1)24.1 (1) Compound 174 not tested 57.0 (1) Compound 175 not tested 5.1(1) Compound 176 not tested 10.0 (1) Compound 177 not tested 0.754 (1)Compound 178 not tested 1.25 (1) Compound 179 not tested 1.67 (1)Compound 180 <100 (1) <10 (1) Compound 181 0.081 (1) 0.632 (1) Compound182 2.6 (1) 7.05 (1) Compound 183 0.676 (1) 5.01 (1) Compound 184 1.5(1) 1.51 (1) Compound 185 0.646 (1) 0.639 (1) Compound 186 0.155 (1)0.123 (1) Compound 187 1.78 (1) 2.01 (1) Compound 188 not tested nottested Compound 189 not tested not tested Compound 190 not tested nottested Compound 191 not tested not tested Compound 192 not tested nottested Compound 193 not tested not tested Compound 194 not tested nottested Compound 195 not tested not tested Compound 196 not tested nottested Compound 197 not tested not tested Compound 198 not tested nottested Compound 199 not tested not tested Compound 200 not tested nottested Compound 201 not tested not tested Compound 202 not tested nottested Compound 203 not tested not tested Compound 204 not tested nottested Compound 205 not tested not tested Compound 206 not tested nottested Compound 207 not tested not tested Compound 208 not tested nottested Compound 209 not tested not tested Compound 210 not tested nottested Compound 211 not tested not tested Compound 212 not tested nottested Compound 213 not tested not tested Compound 214 not tested nottested Compound 215 not tested not tested^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1). (# in parentheses indicates the number of experiments).^(b)TFA salt.^(c)Inhibition of [³H]MK-801 binding in rat cortical/hippocampal washedmembrane preparations (see Example 4).^(d)IC₅₀ study incomplete. % inhibition at the stated concentration.

A comparison of the IC₅₀ values in the RCGC assay with the IC₅₀ valuesin the [³H]MK-801 binding assay (Table 1) illustrates that thearylalkylamines of the invention inhibit NMDA receptor activity by amechanism different than that of binding to the MK-801 binding site; theconcentration of the compound that inhibits NMDA receptor function-isseveral orders of magnitude less than the concentration that competes atthe site labeled by [³H]MK-801. This is not the case, however, with thesimplified arylalkylamines exemplified by Compounds 19-215. Suchcompounds bind to the site labeled by [³H]MK-801 at concentrationsranging approximately 1 to 400-fold higher than those which antagonizeNMDA receptor-mediated function in the rat cerebellar granule cellassay.

Some of the simplified arylalkylamines disclosed have structuralfeatures similar to portions of other compounds which are utilized as,for example, anticholinergics, antiparkinsonians, antihistamines,antidepressants, calcium channel blockers, coronary vasodilators, opiateanalgesics, and antiarrhythmics. However, when certain of thesecompounds were evaluated for NMDA receptor antagonist potency (Example1), as can be seen in Table 2, none of the compounds tested, with theexception of (R)— and (S)-fendiline, nisoxetine, and the Eli Lillycompound, had IC₅₀ values less than 1 μM. These data are summarized inTable 2. TABLE 2 Compound and Therapeutic IC₅₀ (μM ) Utility Structurevs. NMDA^(a) (R)-fendiline (calcium channel blocker; coronaryvasodilator)

0.719 (S)-fendiline (calcium channel blocker; coronary vasodilator)

0.686 prenylamine (calcium channel blocker; coronary vasodilator)

˜10 pheniramine (antihistamine)

22 chlorpheniramine (antihistamine)

>100 brompheniramine (antihistamine)

138 diphenhydramine (antihistamine)

26 doxylamine (antihistamine; hypnotic)

62 chlorcyclizine (antihistamine)

˜10 cyclizine (antiemetic)

28 nor-cyclizine (pharmaceutical intermediate)

23 lidoflazine (calcium channel blocker; coronary vasodilator)

>30 pimozide (antipsychotic)

>10 disopyramide (antiarrhythmic)

>100 isopropamide (antichloinergic)

87 pridinol (anticholinergic; antiparkinsonian)

10.7 chloropyramine (antihistamine)

76 trihexyphenidyl (anticholinergic; antiparkinsonian)

5.13 fluoxetine (antidepressant)

3.4 zimeldine (antidepressant)

≧26 methadone (opiate analgesic)

not tested Astra compound^(b)(antidepressant)

>30 Novo-Nordisk compound^(c)(calcium channel blocker; neuroprotectant)

not tested Novo-Nordisk compound^(d)(calcium channel blocker;neuroprotectant)

28.8 nisoxetine (monoamine uptake inhibitor; antidepressant)

0.894 terodiline (calcium channel blocker; anticholinergic; vasodilator)

not tested tomoxetine (monoamine uptake inhibitor; antidepressant)

not tested amitriptyline (serotonin uptake inhibitor; antidepressant)

not tested imipramine (serotonin uptake inhibitor; antidepressant)

not tested clomipramine (serotonin uptake inhibitor; antidepressant)

not tested doxepine (serotonin uptake inhibitor; antidepressant)

not tested chlorpromazine dopamine otagonist; neuroleptic)

not tested desipramine (antidepressant)

2.3 protriptyline (antidepressant)

≦10 Eli Lilly Compound NMDA receptor antagonist^(e)

0.609

a: Inhibition of NMDA/glycine-induced increases in intracellular calciumin cultured rat cerebellar granule cells (RCGC,s) (see Example 1).

b: Disclosed as compound 2 in Table 4 in Marcusson et al., Inhibition of[³H]paroxetine binding by various serotonin uptake inhibitors:structure-activity relationships. Europ. J. Pharmacol. 215: 191-198,1992.

c: Disclosed as compound 17 in Jakobsen et al.,Aryloxy-phenylpropylamines and their calcium overload blockingcompositions and methods of use. U.S. Pat. No. 5,310,756, May 10, 1994.

d: Disclosed as compound 25 in Jakobsen et al.,Aryloxy-phenylpropylamines and their calcium overload blockingcompositions and methods of use. U.S. Pat. No. 5,310,756, May 10, 1994.

e: Disclosed as Compound 1 in McQuaid et al., Inhibition of [³H]-MK801binding and protection against NMDA-induced lethality in mice by aseries of imipramine analogs Res. Comm. in Pathol. and Pharm.77:171-178, 1992.

Structure-activity relationship studies were initiated using Compound 19as the lead structure. An examination of the side chain demonstratedthat the propyl side chain was optimal for NMDA receptorantagonist-potency (Table 3). This finding was verified using Compound20 as the lead structure (Table 3). TABLE 3 IC₅₀ (μM) vs. CompoundStructure NMDA^(a) 2,2-diphenylethylamine

24.5 3,3-diphenylpropylamine (Compound 19)

0.435 4,4-diphenylbutylamine (Compound 70)

1.7 5,5-diphenylpentylamine (Compound 71)

6.4 2,2-bis(3-fluorophenyl)-1- ethylamine (Compound 98)

7.9 3,3-bis(3-fluorophenyl)-1- propylamine (Compound 20)

0.070 4,4-bis(3-fluorophenyl)-1- butylamine (Compound 100)

0.602^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

Further SAR studies examined the optimal pattern of phenyl ringsubstitution. Initial studies demonstrated that substitution of ahalogen group (fluoro or chloro) at the meta position was optimal forNMDA receptor antagonist potency (Table 4). Increasing the number offluoro substituents led to an apparent decrease in potency (Table 4).TABLE 4 IC₅₀ (μM) vs. Compound Structure NMDA^(a)3,3-diphenyl-1-propylamine (Compound 19)

0.435 3-(2-fluorophenyl)-3-(4- fluorophenyl)-1- propylamine (Compound76)

0.730 3,3-bis(4-fluorophenyl)-1- propylamine (Compound 77)

5.5 3,3-bis(3-fluorophenyl)-1- propylamine (Compound 20)

0.070 3-(2-fluorophenyl)-3-(3- fluorophenyl)-1- propylamine (Compound52)

0.102 3,3-bis(2-fluorophenyl)-1- propylamine (Compound 53)

0.217 3,3-bis(3-chlorophenyl)-1- propylamine (Compound 31)

0.052 3-(3-fluorophenyl)-3-(3- chlorophenyl)-1- propylamine (Compound30)

0.035 3-(3-fluorophenyl)-3- phenyl-1-propylamine (Compound 32)

0.284 3-(3,5-difluorophenyl)-3- (3-fluorophenyl)-1- propylamine(Compound 96)

0.187 3,3- bis(3,5,difluorophenyl)- 1-propylamine (Compound 97)

0.410 3,3-bis[3- (trifluoromethyl)phenyl]- 1-propylamine (Compound 78)

10.2^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

Replacement of one of the fluoro groups on one phenyl ring with amethyl, methoxy or hydroxy group led to no change or a decrease in thein vitro NMDA receptor antagonist potency. The ortho position wasoptimal for this methyl, methoxy or hydroxy group, and the rank order ofpotency for this substitution was methyl>methoxy>hydroxy (Table 5). Alsoillustrated in Table 5 are those compounds possessing the3,3-bis(3-fluorophenyl)moiety with additional methyl or methoxysubstitutions on the phenyl rings, often leading to increase in NMDAreceptor antagonist potency. Table 5 also illustrates those compoundspossessing the 3,3-bis(2-methyphenyl) or 3,3-bis(2-methoxyphenyl)moietyin place of the 3,3-bis(3-fluorophenyl)moiety; these substitutions areacceptable, although a decrease in potency is noted. TABLE 5 IC₅₀ (μM)vs. Compound Structure NMDA^(a) 3,3-bis(3-fluorophenyl)-1- propylamine(Compound 20)

0.070 3-(3-fluorophenyl)-3-(2- methylphenyl)-1- propylamine (Compound27)

0.071 3-(3-fluorophenyl)-3-(3- methylphenyl)-1- propylamine (Compound28)

0.380 3-(3-fluorophenyl)-3-(4- methylphenyl)-1- propylamine (Compound29)

1.9 3-(3-fluorophenyl)-3-(2- methoxyphenyl)-1- propylamine (Compound 24)

0.206 3-(3-fluorophenyl)-3-(3- methoxyphenyl)-1- propylamine (Compound25)

0.279 3-(3-fluorophenyl)-3-(4- methoxyphenyl)-1- propylamine (Compound26)

27 3-(2-methoxyphenyl)-3- phenyl-1-propylamine (Compound 97)

0.410 3-(2-hydroxyphenyl)-3-(3- fluorophenyl)-1- propylamine (Compound103)

0.380 3-(3-hydroxyphenyl)-3-3- fluorophenyl)-1- propylamine (Compound101)

0.912 3-(3-fluorophenyl)-3-(2- methyl-3-fluorophenyl)- 1-propylamine(Compound 56)

0.218 3-(3-fluorophenyl)-3-(3- fluoro-6-methylphenyl)- 1-propylamine(Compound 57)

0.028 3,3-bis(3-fluoro-6- methylphenyl)-1- propylamine (Compound 58)

0.028 3-(3-fluorophenyl)-3-(3- fluoro-6-methoxyphenyl)-1- propylamine(Compound 61)

0.134 3,3-bis(2-methylphenyl)-1- propylamine (Compound 65)

0.167 3,3-bis(2-methoxyphenyl)- 1-propylamine (Compound 62)

0.177 3,3-bis(3-methoxyphenyl)- 1-propylamine (Compound 115)

1.9^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

The next series of SAR experiments investigated the effect of alkylchain substitutions (branching patterns) on NMDA receptor antagonistpotency in vitro. The addition of a methyl group on either the α or βcarbon on the propyl side chain led to a decrease or no change inpotency, respectively (Table 6). TABLE 6 IC₅₀ (μM) vs. CompoundStructure NMDA^(a) 3,3-bis(3-fluorophenyl)-1- propylamine (Compound 20)

0.070 3,3-bis(3-fluorophenyl)-2- methyl-1-propylamine (Compound 21)

0.038 3,3-bis(3-fluorophenyl)-2- methyl-1-propylamine (Compound 33)

0.060 3,3-bis(3-fluorophenyl)-2- methyl-1-propylamine (Compound 34)

0.426 3,3-bis(3-fluorophenyl)-1- methyl-1-propylamine (Compound 22)

0.145 3,2-bis(3-fluorophenyl)-1- methyl-1-propylamine (Compound 50)

0.089 3,3-bis(3-fluorophenyl)-1- methyl-1-propylamine (Compound 51)

1.1 3,3-bis(3-fluorophenyl)-2- ethyl-1-propylamine (Compound 55)

0.035 3,3-bis(3-fluorophenyl)-1- ethyl-1-propylamine (Compound 23)

0.267 3,3-bis(3-fluorophenyl)-2- hydroxyethyl-1-propylamine (Compound54)

0.036 3,3-bis(3-fluorophenyl)-3- ethyl-1-propylamine (Compound 82)

0.106 3,3-bis(3-fluorophenyl)- 1,2-dimethyl-1-propylamine (Compound 38)

0.407 3,3-bis(3-fluorophenyl)- 2,2-dimethyl-1-propylamine (Compound 41)

0.724 3,3-bis(3-fluorophenyl)- 2,2-dimethyl-1-propylamine (Compound 80)

28^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

The next series of SAR experiments investigated the effect ofincorporation of a double bond within the propyl chain on NMDA receptorantagonist potency in vitro (Table 7). As can be seen in Table 7, theincorporation of a double bond decreased potency in a consistent manner.TABLE 7 IC₅₀ (μM) vs. Compound Structure NMDA^(a)3,3-bis(3-fluorophenyl)-1- propylamine (Compound 20)

0.070 3,3-bis(3-fluorophenyl)- prop-2-ene-1-amine (Compound 139)

1.4 3,3-diphenyl-1-propylamine (Compound 19)

0.435 3,3-diphenyl-prop-2-ene-1- amine (Compound 81)

1.4 3-(3-fluorophenyl)-3- phenyl-1-propylamine (Compound 32)

0.284 3-(3-fluorophenyl)-3- phenyl-prop-2-ene-1-amine (Compound 107)

2.67

3,3-bis(3-methoxyphenyl)-1- propylamine (Compound 115)

1.9 3,3-bis(3-methoxyphenyl)- prop-2-ene-1-amine (Compound 116)

4.47^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

The next series of SAR experiments investigated the effect ofincorporation of the propylamine chain into a ring structure on NMDAreceptor antagonist potency in vitro (Table 8) TABLE 8 IC₅₀ (μM) vs.Compound Structure NMDA^(a) 3,3-bis(3-fluorophenyl)-1- propylamine(Compound 20)

0.070 Compound 63

0.093 Compound 64

0.309 Compound 102

1.01 Compound 84

7.9 Compound 111

0.790 Compound 112

28.9^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

The next series of SAR experiments investigated the effect of simplealkyl substitution on the nitrogen on NMDA receptor antagonist potencyin vitro (Table 9) TABLE 9 IC₅₀ (μM) vs. Compound Structure NMDA^(a)3,3-bis(3-fluorophenyl)-1- propylamine (Compound 20)

0.070 N-methyl-3,3-bis(3- fluorophenyl)-1- propylamine (Compound 60)

0.416 N-ethyl-3,3-bis(3- fluorophenyl)-1- propylamine (Compound 59)

0.272 N,N-dimethyl-3,3-bis(3- fluorophenyl)-1- propylamine (Compound123)

9.6 3-(3-fluorophenyl)-3- phenyl-1-propylamine (Compound 32)

0.284 N-methyl-3-(3- fluorophenyl)-3-phenyl)- 1-propylamine (Compound108)

1.06 3,3-diphenylpropylamine (Compound 19)

0.435 N-methyl-3,3- diphenylpropylamine (Compound 67)

10.95 N-ethyl-3,3- diphenylpropylamine (Compound 68)

2.9 N,N-dimethyl-3,3- diphenylpropylamine (Compound 73)

12.6 N-isopropyl-3,3- diphenylpropylamine (Compound 72)

7.4 N,N-diethyl-3,3- diphenylpropylamine (Compound 74)

27.5^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).

Certain simplified arylalkylamine compounds were selected for evaluationof activity in a battery of neurotransmitter receptor binding assays,and for activity against the L-type calcium channel and delayedrectifier potassium channel. The compounds were inactive (less than 50%inhibition at concentrations up to 10 μM) in the following assays:nonselective α2 adrenergic receptor ([³H]RX 821002 binding in ratcortex), H1 histamine receptor ([³H]pyrilamine binding in bovinecerebellum), nonselective sigma receptor ([³H]DTG binding in guinea pigbrain), nonselective opiate receptor ([³H]naloxone binding in ratforebrain), monoamine oxidase (MAO) activity, both MAO-A ([¹⁴C]serotoninmetabolism in rat liver mitochondria) and MAO-B ([¹⁴C]phenylethylaminemetabolism in rat liver mitochondria).

As can be seen in Table 10, activity was noted for several compounds atconcentrations below 10 μM in the following assays: L-type calciumchannel, delayed rectifier potassium channel, central muscariniccholinergic receptor binding, and monoamine (dopamine, norepinephrine,and serotonin) uptake binding assays. This profile of activity in thecentral muscarinic cholinergic receptor and monoamine uptake bindingassays is not unexpected, given the chemical structures of oursimplified arylalkylamines (refer to Table 2 above). With theexceptions, however, of the activity of Compound 19 in the serotoninuptake binding assay, the activity of Compound 34 in the dopamine uptakebinding assay, the activity of Compound 50 in the serotonin uptakebinding assay, the activity of Compounds 63 and 64 in the dopamineuptake binding assay, and the activity of Compound 60 in the dopamineand serotonin uptake binding assays, the simplified arylalkylaminecompounds were most potent at the NMDA receptor. TABLE 10 Central IC₅₀Delayed muscarinic Monoamine (μm) L-type rectifier Choli- uptake vs.calcium potassium nergic binding Compound NMDA^(a) channel^(b)channel^(c) receptor^(d) assays^(e) Compound 0.435 10.2  1-10 4% at 7%at 19 0.174% at 0.1^(f)75% at 10 10^(f)3% at 0.1^(g)53% at 10^(g)18% at0.1^(h)89% at 10^(h) Compound 0.070 2.2 1-10 8% at 6% at 20 0.190% at0.1^(f)81% at 10 10^(f) 5% at 0.1^(g)58% at 10^(g)28% at 0.1^(h)94% at10^(h) Compound 0.00 1.6 >10 42% at 23% at 33 0.199% at 0.1^(f)86% at 1010^(f)2% at 0.1^(G)54% at 10^(g)14% at 0.1^(h)89% at 10^(h) Compound0.426 not ˜10 25% at 60% at 34 tested 0.199% at 0.1^(f)99% at 1010^(f)10% at 0.1^(g)64% at 10^(g)12% at 0.1^(h)79% at 10^(h) Compound0.089 not ˜10 11% at 17% at 50 tested 0.184% at 0.1^(f)93% at 10 10^(f)10% at 0.1^(g)78% at 10^(g)75% at 0.1^(h)97% at 10^(h) Compound 0.0130.676 ˜3 33% at 40% at 46 0.189% at 0.1^(f)97% at 10 10^(f)7% at0.1^(g)64% at 10^(g)10% at 0.1^(h)75% at 10^(h) Compound 0.093 1.9 not11% at 64% at 63 tested 0.181% at 0.1^(f)98% at 10 10^(f)7% at0.1^(g)76% at 10^(g)13% at 0.1^(h)85% at 10^(h) Compound 0.309 not not11% at 50% at 64 tested tested 0.183% at 0.1^(f)99% at 10 10^(f) 8% at0.1^(g)65% at 10^(g)29% at 0.1^(h)68% at 10^(h) Compound 0.028 1.6 not1% at 0% at 58 tested 0.148% at 0.1^(f)45% at 10 10^(f)1% at 0.1^(g)67%at 10^(g)27% at 0.1^(h)95% at 10^(h) Compount 0.272 not not 9% at 2% at59 tested tested 0.187% at 0.1^(f)78% at 10 10^(f)7% at 0.1^(g)51% at10^(g)14% at 0.1^(h)86% at 10^(h) Compound 0.416 2.3 not 13% at 0.91416% 60 tested 0.193% at at 0.1^(g)64% 10 at 10^(g) 0.068^(h)^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's) (see Example1).^(b)Inhibition of KC1 depolarization-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGCs); estimated IC₅₀value in μM.^(c)Inhibition of delayed rectifier potassium channel in culturedNlE-115 neuroblastoma cells; estimated IC₅₀ Value in μM.^(d)Inhibition of the binding of [³H]quinuclidinylbenzilate (QNB) to ratcortical membranes; percent block at indicated concentration in μM.^(e)Inhibition of the binding of [³H]WIN-35,428 to guinea pig striatalmembranes (dopamine uptake binding assay),# [³H]desipramine to rat cortical membranes (norepinephrine uptakebinding assay), or [³H]citalopram to rat forebrain membranes #(serotonin uptake binding assay); percent block at indicatedconcentration in μM, or IC₅₀ when available.^(f)dopamine uptake binding assay^(g)norepinephrine uptake binding assay^(h)serotonin uptake binding assay

Advantageous properties of the arylalkylamine compounds of the presentinvention are illustrated by the fact that concentrations which suppressNMDA receptor-mediated synaptic transmission fail to inhibit LTP.Furthermore, while compounds such as Compound 9, and 11 do produce ahypotensive response following systemic administration in rats, thehypotensive effect produced by these compounds is of a relatively shortduration (approximately 30 min). Additionally, Compounds 12 and 14 haveno cardiovascular activity at doses up to 37.3 μmoles/kg i.v. and 15μmoles/kg i.v., respectively. TABLE 11 Suppresion of NMDA Drop in MeanReceptor-Mediated Arterial Blood Compound Synaptic Transmission^(a) LTPAssay^(b) Pressure^(c) Compound 1  10-30 μM no block at 65 mm Hg at 300μM 1.5 μmoles/kg i.v., 60 min duration Compound 2  10-30 μM no block at40 mm Hg at 100 μM 1.5 μmoles/kg i.v., 120 min duration Compound 3 10-30 μM not tested 20 mm Hg at 1 mg/kg s.c., >60 min duration Compound4  10-100 μM no block at 40 mm Hg at 100 μM 1.5 μmoles/kg i.v., 120 minduration Compound 9  10-100 μM no block at 75 mm Hg at 100 μM 4.5μmoles/kg i.v., 90 min duration Compound 11 not tested not tested 20 mmHg at1 mg/kg i.v., 30 min duration Compound 12 not tested not tested noeffect at doses up to 37.3 μmoles/kg i.v. Compound 14 not tested nottested no effect at doses up to 15 μmoles/kg i.v. Compound 19 100-300 μMbock at 100 μM not tested Compound 20  30-300 μM block at 100 μM noeffect at doses up to 15 μmoles/kg i.v. Compound 22 not tested nottested no effect at doses up to 15 μmoles/kg i.v.^(a)Concentration which suppresses NMDA receptor-mediated synaptictransmission (see Example 5).^(b)Concentration that does not block the induction of LTP (see Example19).^(c)Drop in systemic blood pressure produced by administration ofcompound in rats (see Example 22).Compounds of Formula VIII and Methods of Use

The present invention further provides methods for treating a patienthaving a neurological disease or disorder, comprising administering acompound of Formula VIII:

wherein:

Z is selected from the group consisting of —CH2CH2-, —CH2CH(CH3)-,—CH═CH—, —O—CH2-, —S—CH2-, —O—, and —S—;

X¹ and X² are independently selected from the group consisting of —F,—Cl, —CH3,—OH, and lower O-alkyl in the 1-, 3-, 7-, or 9-substituentpositions.

m is independently an integer from 0 to 2;

—NHR is selected from the group consisting of —NH₂, —NHCH₃, and —NHC₂H₅;

R¹ is selected from the group consisting of —H, alkyl, hydroxyalkyl,—OH, —O-alkyl, and —O-acyl, and

R² is selected from the group consisting of —H, alkyl, hydroxyalkyl, andpharmaceutically acceptable salts and complexes thereof, wherein thecompound is active at an NMDA receptor.

Especially preferred aspects are those embodiments in which:

Z is —CH₂CH₂—;

X¹ or X² is —F, or both X¹ and X² are —F;

either R¹ or R² is methyl or both R¹ and R² are —H; and

—NHR is selected from the group consisting of —NH2 or —NHCH₃.

In further preferred embodiments, the methods of treatment includeadministration of a compound selected from compounds 156, 182, 183, 184,187, 193, 194, 195, 196, and 197, and pharmaceutically acceptable saltaand complexes thereof. Preferably, said compound is active at an NMDAreceptor.

In most preferred embodiments, the methods of treatment includeadministration of a compound selected from compounds 193, 194, 195, 196,and 197.

The present invention further provides simplified arylalkylaminescomprising the compounds of Formula VIII, and all preferred aspects ofFormula VIII, as set out above.

Examples of such simplified arylalkylamines include, but are not limitedto, Compounds 156, 182, 183, 184, 187, 193, 194, 195, 196, and 197,whose structures are provided herein. Preferably, the compound has anIC₅₀ 10 μM at an NMDA receptor. More preferably, the compound has anIC₅₀≦5 μM, more preferably ≦2.5 μM, and most preferably ≦0.5 μM at anNMDA receptor.

In preferred embodiments, the compound is selected from the groupconsisting of Compound 193, 194, 195, 196, and 197, and pharmaceuticallyacceptable salts and complexes thereof.

In most preferred embodiments, the compound, selected from the groupconsisting of Compounds 193-197, is active at an NMDA receptor.

Also provided in an aspect of the invention are pharmaceuticalcompositions of Formula VIII useful for treating a patient having aneurological disease or disorder. The pharmaceutical compositions areprovided in a pharmaceutically acceptable carrier and appropriate dose.The pharmaceutical compositions may be in the form of pharmaceuticallyacceptable salts and complexes, as is known to those skilled in the art.

Preferred pharmaceutical compositions comprise a compound selected fromthe group consisting of Compound 193, 194, 195, 196, 197, andpharmaceutically acceptable salts and complexes thereof, and apharmaceutically acceptable carrier. Preferably, the compound has anIC₅₀≦10 μM at an NMDA receptor. More preferably, the compound has anIC₅₀≦5 μM, more preferably ≦2.5 μM, and most preferably ≦0.5 μM at anNMDA receptor.

Synthetic Scheme for Compound 195

Synthesis of Compound 195 2-Bromomethyl-5-fluorobenzonitrile

A mixture of 5-fluoro-2-methylbenzonitrile (10.0 g, 74.0 mmol),N-bromosuccinimide (NBS, 13.2 g, 74.0 mmol), benzoyl peroxide (0.2 g,0.82 mmol), and carbon tetrachloride (100 mL) was refluxed for 18 h. Thereaction mixture was then cooled to room temperature and filteredthrough fritted glass. The solid in the funnel was washed with carbontetrachloride (3×50 mL) and evaporated to dryness under vacuum toprovide 16.1 g, 102% yield of product.

Diethyl-2-cyano-4-fluorobenzyl phosphonate

A mixture of 2-bromomethyl-5-fluoro-benzonitrile (16.1 g, 75.2 mmol, 1equiv) and triethyl phosphite (16.3 mL, 15.8 g, 94.8 mmol, 1.26 equiv)were heated at 130° C. for 1 h, open to the air. The reaction mixturewas then heated at 165° C. for 30 min to remove ethyl bromide and excesstriethyl phosphite and subsequently cooled to room temperature toprovide 24.8 g (122% crude yield) of a light-orange oil.

2-[2 (4-Fluorophenyl)ethenyl]-5-fluorobenzonitrile

Sodium hydride (60% in mineral oil; 1.66 g [0.993 g NaH], 1.4 mmol, 1.10equiv) was added to a solution of diethyl-2-cyano-4-fluorobenzylphosphonate (10.2 g-equiv, 37.6 mmol, 1.0 equiv) in DMF (40 mL) over aperiod of 2 min. The reaction was stirred for 20 min at roomtemperature. 4-Fluorobenzaldehyde (4.67 g, 38 mmol, 1.0 equiv) was thenadded over a period of 2 min. After stirring for 18 h, H₂O (200 mL) andsatd. NaCl (50 mL) were added and the resulting mixture was extractedwith EtOAc (2×100 mL). The combined organic layers were washed with 1NNaOH (1×100 mL) and H₂O (2×100 mL), dried (anhyd. Na₂SO₄), and rotaryevaporated (>100° C., heat gun). The crude, dark-red crystalline/oil(7.49 g, 82.7%) was chromatographed twice on silica gel (50×250 mm, drypack), eluting first with hexane/EtOAc [4:1] and the secondchromatography with [30:1] to provide a TLC-pure, yellow compound (2.1g, 23.2%).

2-[2-(4-Fluorophenyl)ethyl]-5-fluorobenzonitrile

2-[2-(4-Fluorophenyl)ethenyl]-5-fluorobenzonitrile (2.1 g, 8.7 mmol) wasdissolved in ethylene-glycol dimethyl ether (100 mL). Palladium oncharcoal (10% Pd; 0.20 g) was added, and the reaction mixture was shakenunder 60 psig H₂ for 1 h. The reaction mixture was then filtered throughCelite®, and the filtrate was rotary evaporated (75° C.) to provide 2.1g (99%) of product as a light-yellow oil.

2-[2-(4-Fluorophenyl)ethyl]-5-fluorobenzoic acid

A mixture of 2-[2-(4-fluorophenyl)ethyl]-5-fluorobenzo-nitrile (2.12 g,8.72 mmol, 1 equiv) and aq. NaOH (10N; 3.9 mL, 39.3 mmol, 4.5 equiv) inethylene glycol (25 mL) was heated at 190° C. for 24 h. After-allowingthe mixture to cool, H₂O (100 mL) was added followed by 12N HCl (8 mL).The cloudy mixture precipitated the desired product as a cream-coloredsolid. This material was washed with H₂O (3×20 mL) and dried undervacuum (18 h, 25° C., 0.1 mm Hg) to provide 2.02 g (88.4%) of acream-colored solid.

3,7-Difluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-one

A mixture of 2-[2-(4-fluorophenyl)ethyl]-5-fluorobenzoic acid (2.02 g,7.71 mmol, 1 equiv) and oxalyl chloride (1.37 g, 10.8 mmol, 1.4 equiv)in CH₂Cl₂ (20 mL) was refluxed for 30 min. The resulting solution wasthen added to a stirring suspension of aluminum chloride (1.44 g, 10.8mmol, 1.4 equiv) in CH₂Cl₂ (50 mL). This mixture was refluxed for 1 h.The reaction mixture was then washed with 1N HCL (3×20 mL), aq. satd.NaCl (1×50 mL), and H₂O (1×50 mL). The organic layer was dried (anhyd.Na₂SO₄) and rotary evaporated to provide 1.6 g of a light-orange-coloredsemisolid. This material was chromatographed (3:1 hexane/CHCl₃) throughsilica gel (150 mm×25-mm dia.). Evaporation of the combined fractionscontaining product provided 0.93 g (49%) of the desired product as alight-yellow shiny solid.

3,7-Difluoro-10,11-dihydro-5H-dibenzo[a,d)cyclohepten-5-ylidenenitrile

Diethylcyanomethylphosphonate (1.01 g, 5.72 mmol, 1.50 equiv) wasdissolved in DMF (15 mL) and sodium hydride (60% in mineral oil; 0.229g, 0.137 g NaH, 5.72 mmol, 1.5 equiv) was added. After stirring for 20min, a solution of 3,7-difluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-one (0.930 g, 3.81 mmol, 1 equiv) in DMF (5 mL) was-added.After stirring for 2 h, EtOAc (100 mL) was added, and the resultingsolution was washed with H₂O. (4×25 mL), dried (anhyd. Na₂SO₄), androtary evaporated (75° C.) to provide 0.86 g (84.5%) of a yellow,crystalline solid.

3,7-Difluoro-10,11-dihydro-5-cyanomethyl-5H-dibenzo [a,d]cycloheptene

Aluminum/mercury amalgam was prepared as follows. To aluminum granules(4.0 g, Aldrich, −10+60 mesh, 99+%) was added 0.5N NaOH (200 mL). Themixture was stirred for 5 min, and then the supernatant was decanted.The etched Al was subsequently washed with H₂O (200 mL), and then withEtOH (200 mL). The aluminum was then suspended in EtOH (100 mL) and asolution of Hg₂Cl (4.0 g, Aldrich, 99.5%, A.C.S. reagent) dissolved inEt₂O (200 mL) was added. The reaction was stirred for 5 min. Thesupernatant was decanted and the Al(Hg) amalgam was washed with H₂O (200mL)), EtOH (200 mL), and, finally, Et₂O (200 mL). The amalgam was thencovered with Et₂O (100 mL) and was ready for use.

A solution of3,7-difluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-ylidenenitrile(0.86 g, 3.22 mmol) dissolved in EtOAc (25 mL) was added to the amalgam.EtOH (25 mL) and then H₂O (5 mL) were then added, and the mixture wasstirred at room temperature for 1 h. The reaction mixture wassubsequently filtered through paper and the filtrate was evaporated toprovide 960 mg (110% yield) of crude product as a nearly colorless oilwhich crystallized on standing. This material was found to becontaminated with a small amount (<5%) of diethylcyanomethylphosphonate.

3,7-Difluoro-10,11,-dihydro-5-(2-aminoethyl)-5H-dibenzo[a,d]cycloheptenehydrochloride

To a suspension of lithium aluminum hydride (339 mg, 8.92 mmol, 2.5equiv) in anhyd. diethyl ether (10 mL) was added3,7-difluoro-10,11-dihydro-5-(2-cyanoethyl)-5H-dibenzo[a,d]cycloheptene(0.96 g, 3.57 mmol) dissolved in anhyd. diethyl ether (5 mL) over aperiod of 2 min. The reaction was then stirred at room temperature for30 min. Water (0.3 mL) was added followed by 5N NaOH (0.3 mL), followedby H₂O (0.9 mL) with stirring to precipitate the inorganic salts.Diethyl ether (20 mL) was added and the reaction was filtered throughpaper. The filtrate was-then rotary-evaporated to provide 0.80 g (82.1%)of product as its free base, a colorless oil which solidified onstanding. The free base was dissolved in diethyl ether (50 mL). Excessethereal hydrogen chloride (1 M) was added to precipitate thecorresponding hydrochloride salt as a white solid (0-60 g, 54%).

Synthetic Scheme for Compound 197

EXAMPLE 2 Synthesis of Compound 1972-[2-(4-Fluorophenyl)ethenyl]benzonitrile

A mixture of (cc-bromo-o-tolunitrile (10.0 g, 51.0 mmol, 1 equiv) andtriethyl phosphite (11.0 mL, 10.7 g, 64.1 mmol, 1.26 equiv) was heatedto 130° C. in an oil bath for 1 h. The reaction was then heated to 165°C. for 30 min to remove ethyl bromide and excess triethyl phosphite.After allowing the reaction to cool, anh. DMF (40 mL) was added. Sodiumhydride (60% dispersion in mineral oil; 2.05 g, 1.23 g NaH, 51.3 mmol,1.00 equiv) was added over a period of 2 min. The reaction was stirredfor 10 min.

The reaction was then placed in a water bath (25° C.) and4-fluorobenzaldehyde (5.5 mL, 6.4 g, 51 mmol, 1.0 equiv) was added overa period of 2 min. After stirring for 2.5 h, H₂O (200 mL) was added andthe resulting mixture was extracted with EtOAc (2×100 mL). The combinedorganic layers were washed with 1N NaOH (1×100 mL) and H₂O (2×100 mL),dried (anh. Na₂SO₄), and rotary evaporated (>100° C., heat gun). Whilestill hot, hexane (25 mL) was added. The product crystallized whilestirring as the mixture was allowed to cool to 25° C. The crystals werefiltered, washed with hexane (2×25 mL), and allowed to dry in the openair for 16 h. This provided 8.10 g (71.1%) of the desired product as anorange, crystalline solid.

2-[2-(4-Fluorophenyl)ethyl]benzonitrile

2-[2-(4-Fluorophenyl)ethyl]benzonitrile (8.10 g, 36.3 mmol) wasdissolved in ethylene glycol dimethyl ether (200 mL). Palladium oncharcoal (10% Pcl; 0.81 g) in H₂O (1.0 mL) was added, and the reactionmixture was shaken under 60 psig H₂ for 1 h. The reaction mixture wasthen filtered through Celite®, and the filtrate was rotary evaporated(75° C.). This provided 8.03 g (98.3%) of the product as an orange oilwhich crystallized on standing.

2-[2-(4-Fluorophenyl)ethyl]benzoic acid

A mixture of 2-[2-(4-fluorophenyl)ethyl]benzonitrile (8.03 g, 35.6 mmol,1 equiv) and aq. NaOH (10N; 16 mL, 160 mmol, 4.5 equiv) in ethyleneglycol (100 mL) was heated to 190 C for 69 h. After allowing thereaction mixture to cool, H₂O (300 mL) was added, followed by 12N HCl(16 mL). This mixture was extracted with CHCl₃ (1×100 mL) and theorganic layer was then washed with H₂O (2×50 mL). The organic layer wasdried (anh. Na2SO4) and rotary evaporated (90° C.). This provided 7.44 g(85.4%) of a light-yellow solid.

3-Fluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-one

A mixture of 2-[2-(4-fluorophenyl)ethyl]benzoic acid (4.00 g, 16.4 mmol,1 equiv) and oxalyl chloride (2.0 mL, 2.9 g, 23 mmol, 1.4 equiv) inCH₂Cl₂ (50 mL) was refluxed for 30 min. The resulting solution was thenadded to a stirring suspension of aluminum chloride (3.0 g, 2.2 mmol,1.4 equiv) in CH₂Cl₂ (50 mL). This mixture was refluxed for 1 h. Thereaction mixture was then washed with 1N HCl (3×50 mL), aq. satd. NaCl(1×50 mL), and H₂O (1×50 mL). The organic layer was dried (anh. Na₂SO₄)and rotary evaporated (75° C.) to provide 3.7 g of a dark-brown oil.This oil was flash chromatographed (20:1 hexane/EtOAc) through flashsilica gel (250 mm×25 mm). Evaporation of the combined fractions ofinterest provided 1.58 g (42.6%) of the desired product as a yellow oil.

3-Fluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-ylidenenitrile

Diethylcyanomethylphosphonate (1.76 g, 9.94 mmol, 1.50 equiv) wasdissolved in DMF (15 mL) and sodium hydride (60% in mineral oil; 0.40 g,0.24 g NaH, 10 mmol, 1.5 equiv) was added. After stirring for 10 min,3-fluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-one (1.50 g, 6.63mmol, 1 equiv) was transferred in with DMF (5 ML). After stirring for 19h, H₂O (100 mL) was added. The resulting mixture was extracted withEtOAc (2×50 mL). The combined organic layers were washed with aq. satd.NaCl (2×50 mL), dried (anh. Na₂SO₄), and rotary evaporated (75° C.) toprovide 2 g of a dark-brown oil. This material was flash chromatographed(9:1 hexane/EtOAc) through flash silica gel (200 mm×25 mm) to provide1.60 g (96.8%) of a yellow solid.

3-Fluoro-10,11-dihydro-5-(2-aminoethyl)-5H-dibenzo[a,d]cycloheptenehydrochloride

3-Fluoro-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-ylidenenitrile (1.60g, 6.42 mmol) was dissolved in EtOH (160 mL), and 1N NaOH (16 mL) andRaney nickel (slurry in H₂O; 1.6 g) were added. The reaction mixture wasshaken under 60 psig H₂ at 60° C. for 16 h. At this point, all of thestarting material had disappeared, but a significant amount ofalkeneamine intermediate remained. The reaction mixture was filteredthrough Celite® and the solvent was removed by rotary evaporation. Theresulting material was dissolved in EtOAc (100 mL), washed with H₂O(2×50 mL), dried (anh. Na₂SO₄), and the EtOAc was removed by rotaryevaporation. This material was then dissolved in EtOH (50 mL). Palladiumon charcoal (10% Pd; 0.80 g) and 12.1N HCl (˜3 mL) were subsequentlyadded. The reaction mixture was then shaken under 60 psig H₂ at 70° C.for 24 h. The reaction mixture was filtered through Celite® and thefiltrate was rotary evaporated to afford 2.10 g of product as a brown,bubbly solid. This solid was dissolved in CHCl₃ (50 mL) and “free-based”with satd. aq. NaHCO₃ (25 mL). The aqueous layer (which was full ofprecipitate)-was then extracted with CHCl₃ (1×50 mL). The combinedorganic layers were dried (anh. Na₂SO₄) and rotary evaporated (75° C.).The resulting dark-brown oil was flash chromatographed (1:10 MeOH/CHCl₃)through flash silica gel (250 mm×25 mm). This yielded 0.48 g offree-base product as a light-brown oil. This oil was dissolved in EtOH(2.0 mL), and 1.0 M HCl in Et20 (3.0 mL) was added. This solution wasrotary evaporated to yield 0.55 g (29%) of the desired product as acream-colored, bubbly solid.

Different possible “planar” ring conformations of Compound 20, andCompounds 194 and 195 are presented below. All are considered to bewithin the scope of the present invention.

Physical Data for Compounds 93-97

Gas chromatographic and mass spectral data were obtained using aHewlett-Packard 5890 Series II Gas Chromatograph with a 5971 Series MassSelective Detector [Ultra-2 Ultra Performance Capillary Column(crosslinked 5% phenyl methyl silicone); column length, 25 m, columni.d., 0.20 mm; He flow-rate, 60 mL/min; injector temp., 250° C.;gradient temperature program, 20° C./min from 125 to 325° C. for 10 min,then held constant at 325° C. for 6 min].

Compound 193, R_(t)=8.79 min, m/z (% rel. int.) 273 (M+,21), 256 (53),255 (32), 242 (13), 241 (73), 231 (11), 230 (75), 229 (100), 228 (29),227 (56), 225 (11), 220 (8), 215 (31), 214 (67), 209 (13), 207 (20), 201(19), 196 (11), 183 (12), 147 (13), 146 (10), 134 (16), 133 (77), 109(42), 107 (7), 83 (8), 44 (44), 43 (8), 42 (8).

Compound 194, R_(t)=8.79 min, m/z (% rel. int.) 273 (M+,22), 256 (59),255 (36), 242 (17), 241 (92), 230 (54), 229 (90), 228 (41), 227 (76),225 (15), 221 (8), 220 (10), 215 (34), 214 (67), 209 (16), 207 (25), 201(27), 19, (12), 183 (15), 147 (14), 146 (13), 134 (20), 133 (100), 109(53), 107 (10), 83 (12), 44 (47), 43 (10), 42 (10).

Compound 195, R_(t)=8.84 min, m/z (% rel. int.) 273 (M+,35), 256 (71),255 (45), 242 (19), 241 (100), 240 (9), 230 (45), 229 (83), 228 (40),227 (77), 225 (15), 220 (8), 215 (31), 214 (56), 209(14), 207 (18), 201(22), 196 (10), 183 (12), 147 (16), 146 (11), 134 (18), 133 (87), 109(47), 107 (8), 83 (10), 44 (45), 43 (9), 42 (9).

Compound 196, R_(t)=8.77 min, m/z (% rel. int.) 255 (M+,13), 238 (43),237 (30), 224 (11), 223 (57), 220 (7), 212 (42), 211 (100), 210 (31),209 (52), 208 (8), 207 (20), 197 (23), 196 (67), 191 (10), 189 (16), 183(30), 178 (7), 170 (9), 165 (8), 134 (9), 133 (40), 116 (7), 115 (39),109 (15), 91 (17), 63 (7), 44 (14), 42 (7).

Compound 197, R_(t)=8.77 min, m/z (% rel. int.) 255 (M+,17), 237 (39),224 (14), 223 (73), 220 (17), 219 (10), 212 (36), 211 (100), 210 (38),209 (64), 208 (11), 207 (25), 205 (12), 197 (27), 196 (71), 194 (12),193 (24), 191 (20), 189 (20), 183 (34), 178 (18), 170 (10), 165 (13),133 (49), 116 (10), 115 (60), 109 (19), 91 (24), 44 (19).

Some of the simplified arylalkyl amines disclosed have structuralfeatures similar to portions of the following compounds.

Biological data for certain tricyclic compounds of Formula VIII areprovided in Table 12. The data presented in Table 12 was obtainedaccording to the same methods used to obtain the data for Table 1. TABLE12 Biological Data for Compounds Claimed IC₅₀ (μM) IC₅₀ (μM) Compoundvs. NMDA^(a) vs. [³H]MK-801^(b) Compound 156 0.069 (3) 0.081 (3)Compound 182 2.6 (1) 7.05 (1) Compound 183 0.676 (1) 5.01 (1) Compound184 1.5 (1) 1.65 (2) Compound 187 1.78 (1) 2.06 (2) Compound 193 nottested inactive @ 1 μM Compound 194 not tested 0.371 (2) Compound 195not tested 0.029 (2) Compound 196 not tested 2.24 (1) Compound 197 nottested 0.053 (2) Eli Lilly Compd. 0.609 (2) 1.17 (3) Amitriptyline nottested 14.2 (2) Imipramine not tested 6.75 (2) Clomipramine not tested10.0 (2) Doxepin not tested 30.4 (1) Chlorpromazine not tested 11.2 (1)Desipramine 2.3 3.26 (2) Protriptyline 73% @ 10 μM 21.4 (1)Nortriptyline 3.7 5.67 (1) Maprotiline 5.8 not tested Cyclobenzaprinenot tested 14.5 (1)^(a)Inhibition of NMDA/glycine-induced increases in intracellularcalcium in cultured rat cerebellar granule cells (RCGC's). (# inparentheses indicates the number of experiments).^(b)Inhibition of [3H]MK-801 binding in rat cortical/hippocampal washedmembrane preparations.

Formulation and Administration

As demonstrated herein, useful compounds of this invention and theirpharmaceutically acceptable salts may be used to treat neurologicaldisorders or diseases. While these compounds will typically be used intherapy for human patients, they may also be used to treat similar oridentical diseases in other vertebrates such as other primates, farmanimals such as swine, cattle and poultry, and sports animals and petssuch as horses, dogs and cats.

In therapeutic and/or diagnostic applications, the compounds of theinvention can be formulated for a variety of modes of administration,including systemic and topical or localized administration. Techniquesand formulations generally may be found in Remington's PharmaceuticalSciences, Mack Publishing Co., Easton Pa. (18th ed. 1990).

Pharmaceutically acceptable salts are generally well known to those ofordinary skill in the art, and may include, by way of example but notlimitation, acetate, benzenesulfonate, besylate, benzoate, bicarbonate,bitartrate, calcium edetate, camsylate, carbonate, citrate, edetate,edisylate, estolate, esylate, fumarate, gluceptate, gluconate,glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate,lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate,napsylate, nitrate, pamoate (embonate), pantothenate,phosphate/disphosphate, polygalacturonate, salicylate, stearate,subacetate, succinate, sulfate, tannate, tartrate, or teoclate. Otherpharmaceutically acceptable salts may be found in, for example,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.(18th ed. 1990).

Preferred pharmaceutically acceptable salts include, for example,acetate, benzoate, bromide, carbonate, citrate, gluconate, hydrobromide,hydrochloride, maleate, mesylate, napsylate pamoate (embonate),phosphate, salicylate, succinate, sulfate, or tartrate.

The useful compounds of this invention may also be in the form ofpharmaceutically acceptable complexes. Pharmaceutically acceptablecomplexes are known to those of ordinary skill in the art and include,by way of example but not limitation, 8-chlorotheophyllinate (teoclate).

The exact formulation, route of administration and dosage can be chosenby the individual physician in view of the patient's condition. (Seee.g. Fingl et al., in The Pharmacological Basis of Therapeutics, 1975,Ch. 1 p. 1).

It should be noted that the attending physician would know how and whento terminate, interrupt, or adjust administration due to toxicity ororgan dysfunction. Conversely, the attending physician would also knowto adjust treatment to higher levels if the clinical responses were notadequate (precluding toxicity). The magnitude of an administered dose inthe management of the disorder of interest will vary with the severityof the condition to be treated and to the route of administration. Theseverity of the condition may, for example, be evaluated in part, bystandard prognostic evaluation methods. Further, the dose and perhapsdose frequency, will also vary according to the age, body weight, andresponse of the individual patient. A program comparable to thatdiscussed above may be used in veterinary medicine.

Depending on the specific conditions being treated, such agents may beformulated into liquid or solid dosage forms and administeredsystemically or locally. The agents may be delivered, for example, in atimed or sustained-release form as is known to those skilled in the art.Techniques for formulation and administration may be found inRemington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.(18th ed. 1990). Suitable routes may include oral, buccal, sublingual,rectal, transdermal, vaginal, transmucosal, nasal or intestinaladministration; parenteral delivery, including intramuscular,subcutaneous, intramedullary injections, as well as intrathecal, directintraventricular, intravenous, intraperitoneal, intranasal, orintraocular injections, just to name a few.

For injection, the agents of the invention may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHank's solution, Ringer's solution, or physiological saline buffer. Forsuch transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants aregenerally known in the art.

Use of pharmaceutically acceptable carriers to formulate the compoundsherein disclosed for the practice of the invention into dosages suitablefor systemic administration is within the scope of the invention. Withproper choice of carrier and suitable manufacturing practice, thecompositions of the present invention, in particular, those formulatedas solutions, may be administered parenterally, such as by intravenousinjection. The compounds can be formulated readilyusing-pharmaceutically acceptable carriers well known in the art intodosages suitable for oral administration. Such carriers enable thecompounds of the invention to be formulated as tablets, pills, capsules,liquids, gels, syrups, slurries, suspensions and the like, for oralingestion by a patient to be treated.

Agents intended to be administered intracellularly may be administeredusing techniques well known to those of ordinary skill in the art. Forexample, such agents may be encapsulated into liposomes, thenadministered as described above. Liposomes are spherical lipid bilayerswith aqueous-interiors. All molecules present in an aqueous solution atthe time of liposome formation are incorporated into the aqueousinterior. The liposomal contents are both protected from the externalmicroenvironment and, because liposomes fuse with cell membranes, areefficiently delivered into the cell cytoplasm. Additionally, due totheir hydrophobicity, small organic molecules may be directlyadministered intracellularly.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve its intended purpose. Determination of theeffective amounts is well within the capability of those skilled in theart, especially in light of the detailed disclosure provided herein.

In addition to the active ingredients, these pharmaceutical compositionsmay contain suitable pharmaceutically acceptable carriers comprisingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Thepreparations formulated for oral administration may be in the form oftablets, dragees, capsules, or solutions.

The pharmaceutical compositions of the present invention may bemanufactured in a manner that is itself known, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspension. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidester, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as-sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

Pharmaceutical preparations for oral use can be obtained by combiningthe active compounds with solid excipients, optionally grinding aresulting mixture, and processing the mixture of granules, after addingsuitable auxiliaries, if desired, to obtain tablets or dragee cores.Suitable excipients are, in particular, fillers such as sugars,including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations, for example, maize starch, wheat starch, rice starch,potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose (CMC),and/or polyvinylpyrrolidone (PVP: povidone). If desired, disintegratingagents may be added, such as the cross-linked-polyvinylpyrrolidone,agar, or alginic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethyleneglycol (PEG), and/or titanium dioxide, lacquer solutions, and suitableorganic solvents or solvent mixtures. Dye-stuffs or pigments may beadded to the tablets or dragee coatings for identification or tocharacterize different combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin, and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols (PEGs). In addition, stabilizers may be added.

Other embodiments are within the following claims.

1. (canceled)
 2. A compound of the formula:

wherein: X is independently selected from the group consisting of: —H,—Br, —Cl, —F, —I, —CF₃, alkyl, —OH, —OCF₃, —O-alkyl, and —O-acyl; R₁ isindependently selected from the group consisting of: —H, lower alkyl,and —O-acyl; R₂ is independently selected from the group consisting of:—H, alkyl, and hydroxyalkyl, or both R₂s together are imino; R₄ isphenoxy which is optionally substituted with —F, —Cl, —Br, —I, —CF₃,alkyl, —OH, —OCF₃, —O-alkyl, or —O-acyl; provided that when X is H, thenR₄ is substituted with at least one of: meta-fluoro, meta-chloro,meta-methyl, meta-OH, ortho-fluoro, ortho-chloro, ortho-methyl, orortho-OH; m is independently an integer from 1 to 5; andpharmaceutically acceptable salts and complexes thereof.
 3. A compoundof the formula:

wherein: X is independently selected from the group consisting of: —H,—Br, —Cl, —F, —I, —CF₃, alkyl, —OH, —OCF₃, —O-alkyl, and —O-acyl; R₁ isindependently selected from the group consisting of: —H, lower alkyl,and —O-acyl; R₂ is independently selected from the group consisting of:—H, alkyl, and hydroxyalkyl, or both R₂s together are imino; R₄ isphenoxy which is optionally substituted with —F, —Cl, —Br, —I, —CF₃,alkyl, —OH, —OCF₃, —O-alkyl, or —O-acyl; provided that when X is H, thenR4 is substituted with at least one of: meta-fluoro, meta-chloro,meta-methyl, meta-OH, ortho-fluoro, ortho-chloro, or ortho-OH; m isindependently an integer from 1 to 5; and pharmaceutically acceptablesalts and complexes thereof.
 4. A compound of the formula:

wherein: X is —H; R₁ is independently selected from the group consistingof: —H, lower alkyl, and —O-acyl; R₂ is independently selected from thegroup consisting of: —H, alkyl, and hydroxyalkyl, or both R₂s togetherare imino; R₄ is phenoxy which is substituted with at least one of —F,—Br, —I, alkyl, —OH, —OCF₃, or —O-acyl; and pharmaceutically acceptablesalts and complexes thereof.
 5. A pharmaceutical composition, comprisinga compound as in any of claims 2-4, and a pharmaceutically acceptablecarrier.
 6. The pharmaceutical composition of claim 5, wherein saidpharmaceutical composition is adapted for the treatment of aneurological disease or disorder.
 7. The pharmaceutical composition ofclaim 5, wherein said pharmaceutical composition is adapted to provideneuroprotection to a patient.
 8. The pharmaceutical composition of claim5, wherein said compound is a hydrochloride salt.
 9. A method fortreating a patient having a neurological disease or disorder, comprisingadministering a compound as in any of claims 2-4.
 10. The method ofclaim 9, wherein said neurological disease or disorder is selected fromthe group consisting of stroke, head trauma, spinal cord injury,epilepsy, anxiety, Alzheimer's disease, Huntington's disease,Parkinson's disease, and arnyotrophic lateral sclerosis.
 11. The methodof claim 10, wherein said neurological disease or disorder is stroke.12. The method of claim 10, wherein said neurological disease ordisorder is head trauma.
 13. The method of claim 10, wherein saidneurological disease or disorder is spinal cord injury.
 14. The methodof claim 10, wherein said neurological disease or disorder is epilepsy.15. The method of claim 10, wherein said neurological disease ordisorder is anxiety.
 16. The method of claim 10, wherein saidneurological disease or disorder is Alzheimer's disease.
 17. The methodof claim 10, wherein said neurological disease or disorder isHuntington's disease.
 18. The method of claim 10, wherein saidneurological disease or disorder is Parkinson's disease.
 19. The methodof claim 10, wherein said neurological disease or disorder isamyotrophic lateral sclerosis.
 20. The method of claim 11, wherein saidstroke is global ischemic.
 21. The method of claim 11, wherein saidstroke is hemorrhagic.
 22. The method of claim 11, wherein said strokeis focal ischemic.
 23. A method for providing neuroprotection to apatient, comprising administering a compound as in any of claims 2-4.